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  • 1985-1989  (356)
  • 1955-1959
  • 1930-1934
  • 1890-1899
  • Immunocytochemistry
  • 1
    Digitale Medien
    Digitale Medien
    Springer
    Virchows Archiv 415 (1989), S. 259-263 
    ISSN: 1432-2307
    Schlagwort(e): Transthyretin ; Human ; Fetus ; In situ hybridization ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Five human fetuses at mid-term (16–20 weeks) and one with a gestational age of 8 weeks were investigated. The cellular localization of transthyretin (TTR)-mRNA in different organs was demonstrated by in situ hybridization with a35S-labelled, single-stranded RNA probe. Immunoreactive TTR (TTR-IR) was localized with a monoclonal antibody to TTR. In all fetuses, choroid plexus epithelial cells demonstrated intense labelling for TTR-mRNA as well as strong TTR-IR. Hepatocytes, on the other hand, showed weak in situ labelling and weak or, in some cases, non-demonstrable TTR-IR. In all mid-term fetuses, but not in the 8 week fetus, TTR-mRNA and TTR were also expressed in pancreatic endocrine A-cells. The degree of in situ labelling in these A-cells was moderate, whereas that of TTR-IR was strong. Despite negative findings for TTR-mRNA in the gut and the kidney, endocrine cells of the gut and epithelial cells of the renal proximal convoluted tubules showed TTR-IR in some of the fetuses. The investigation provides evidence for TTR synthesis in the human fetal choroid plexus, liver and endocrine pancreas. However, further studies are required to demonstrate TTR synthesis in the gut and the kidney.
    Materialart: Digitale Medien
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  • 2
    ISSN: 1432-2307
    Schlagwort(e): Bone Gla protein ; Osteocalcin ; Immunocytochemistry ; Calcification ; Osteogenesis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Boneγ-carboxyglutamic acid containing protein (BGP) was isolated from human bone and anti-BGP antibody was produced in rabbits. Localization of BGP was investigated immunocytochemically by light and electron microscopy in human bones in various developmental stages and pathological conditions. In the bones of a 12 week fetus, osteoblasts stained strongly in areas of bone formation. However in bones of late fetal stages and in newborns and adults, BGP was localized predominantly in the osteoid and bone matrix in the ossifying front. Osteoblasts and osteocytes also stained positive, but less dominantly than in the early fetus. Electron microscopy showed that BGP was localized in the ER and Golgi cisternae of osteoblasts and osteocytes, and the collagen fibers of the osteoid and bone matrix. The intensity and distribution of staining were not significantly different in osteoporosis and osteoarthritis. These observations indicate that BGP is synthesized by osteoblasts most actively in early fetal life and is then deposited on collagen fibers of the osteoid and bone matrix.
    Materialart: Digitale Medien
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  • 3
    ISSN: 1432-2307
    Schlagwort(e): Metallothionein ; Immunocytochemistry ; Embryonal carcinoma
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary The presence of high levels of metallothionein (MT) in developing mammalian cells is well documented. It has been suggested that the developmental profile and gene expression of MT is similar to that of the so-called oncodevelopmental gene products such as a-fetoprotein. In this study tissue sections of nine human embryonal carcinomas of the testis were tested by means of the avidin-biotin peroxidase complex for the presence of MT. The antigen was localized in variable amounts in the cytoplasm and nucleus in tumour cells in all cases. There was evidence that immunoreactivity was related to the histological growth pattern of tumour cells. These findings suggest that MT may be considered an oncodevelopmental product which could be useful as a tumour marker. In addition, the histology of these tumours might predict MT expression; this may prove of value in testing the hypothesis of MT-related emergence of drug-resistant cell lines in the course of treatment of tumours with metal-containing chemotherapeutic agents.
    Materialart: Digitale Medien
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  • 4
    ISSN: 1432-2307
    Schlagwort(e): Bone tumors ; Osteosarcoma ; Osteonectin ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary 18 bone-forming tumours and tumour-like lesions were investigated immunocytochemically for the presence of osteonectin. A group of non-bone-forming skeletal tumours (five cartilage-forming tumours, four Ewing sarcomas and five extraskeletal sarcomas) served as controls. The studies showed that osteonectin antibodies react reliably with benign and malignant bone-forming tumours (two cases of fibrous dysplasia, three osteoid osteomas, 13 osteosarcomas). This finding was supported by protein blot studies. Osteonectin is formed by cells which do not yet possess the morphological phenotype of osteoblasts and may be regarded as a “differentiation marker” of the osteoblastic lineage. Only chondroid bone (tissue in which chondrocytes were surrounded by osteoid matrix containing type I and type II collagen) showed a positive reaction. All other primary skeletal tumours and extraskeletal soft tissue tumours were completely negative.
    Materialart: Digitale Medien
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  • 5
    ISSN: 1432-2307
    Schlagwort(e): IGF-II ; Carotid body Extra-adrenal paraganglioma ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Insulin-like-growth factor (IGF)-II-like immunoreactivity was examined in two carotid bodies and six extra-adrenal paragangliomas with use of monoclonal antibody against rat IGF-II, which crossreacts with human IGF-II. Chief cells but not sustentacular cells of the carotid body were positive at about 10% in one case and less than 1% in another case. Among four carotid body tumours, a possible vagal body tumour and one glomus jugulare tumour, all but the glomus jugulare tumour exhibited positive tumour cells irrespective of histological variations. The frequency of positive cells ranged from 20 to 60%. IGF-II like immunoreactivity, therefore, might be widely distributed in human extra-adrenal paraganglionic tissues and tumours, although its biological role in these cells remains to be elucidated.
    Materialart: Digitale Medien
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  • 6
    ISSN: 1432-2307
    Schlagwort(e): Human acute pancreatitis ; Zymogen granules ; Acinar cells ; Electron microscopy ; Immunocytochemistry ; Morphometry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary In the present study fine structural changes of acinar zymogen granules were investigated in human acute pancreatitis. Pancreatic tissue was obtained at surgery from 6 patients, prepared for ultrastructural analysis, and stained immunocytochemically for trypsinogen. Stereological parameters of zymogen granules were evaluated. The density of the immunocytochemical labelling for trypsinogen was estimated over zymogen granules, the rough endoplasmic reticulum, Golgi apparatus and the acinar lumina. In acute pancreatitis the number of zymogen granules was diminished and their size reduced. The density of the labelling for trypsinogen was unchanged over zymogen granules but showed a significant reduction over the rough endoplasmic reticulum, Golgi apparatus, and the acinar lumina. In general the integrity of zymogen granules was well preserved. Focally degenerative changes of zymogen granules and large autophagosomes were observed. From the immunogold labelling a disturbance of enzyme synthesis and secretion was suggested. Evidence is given that a disruption of the zymogen granule membranes and a fusion with lysosomal bodies might contribute to the pathogenesis of human acute pancreatitis.
    Materialart: Digitale Medien
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  • 7
    ISSN: 1432-2307
    Schlagwort(e): C-peptide ; Insulin ; Pancreaticβ cells ; Insulinoma morphometry ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary It has been shown, by using the immunogold technique, that C-peptide and insulin are co-localized in the mature granules of human pancreaticβ cells and insulinomas with typical granules. The mean gold bead densities of both C-peptide and insulin were at least twice as high in the normal pancreas when compared with the insulinomas. The mean granule diameter of the insulinoma cells (D=0.30 ±0.12 μm) was smaller than that of human pancreatic cells (D=0.45 ±0.15 μm). The morphometric data indicate that each of the antigens (C-peptide and insulin) is distributed similarly in the halos and the dense cores of theβ granules. Thus, no topological segregation of these two antigens occurs within theβ granules of either normal human pancreas or insulinomas.
    Materialart: Digitale Medien
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  • 8
    ISSN: 1432-2307
    Schlagwort(e): Cystic fibrosis ; Endocrine pancreas ; Immunocytochemistry ; Morphometry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary The pancreases of 23 patients (mean age 10.5 years, range 5-22) years dying of cystic fibrosis (CF) were evaluated at autopsy by routine histology and immunostaining for changes in their endocrine cell compartment. The severely altered pancreatic tissues showed end stage CF, with either a fibrotic pattern (CF-FIB,n=14) or a lipoatrophic pattern (CF-LIP,n=9) prevailing. In all specimens, irrespective of the dominating pattern, the islet system was affected by marked periinsular and intrainsular sclerosis. Quantitatively, the volume densities (relative tissue components) of the parenchymal, fibrotic, fatty and total endocrine compartments as well as the four islet cell types (B, A, D, PP) were determined by point counting. Compared with controls, the CF patients (including two patients with overt diabetes and glucose intolerance, respectively) had a significantly decreased insulin (B)-cell ratio (from 64.4 to 34%) with a concomitant rise in non-B-cells (A-cells: 23.2 to 35%; D-cells: 10.4 to 22%; PP-cells; 2 to 9%). Comparison of endocrine cell ratios in CF-FIB pancreases with CF-LIP pancreases revealed no significant differences. The reduction of approximately 50% of insulin cells in CF patients with advanced disease supports the concept that destruction of exocrine tissue with concomitant fibrous disorganization of islets gradually changes the proportional distribution of the endocrine cells in favor of the noninsulin cells. This slowly ongoing process probably provides the basis for islet dysfunction, i.e. diabetes, increasingly observed in final stage CF.
    Materialart: Digitale Medien
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  • 9
    Digitale Medien
    Digitale Medien
    Springer
    Virchows Archiv 416 (1989), S. 81-89 
    ISSN: 1432-2307
    Schlagwort(e): Gangliocytic paraganglioma ; Duodenum ; Immunocytochemistry ; Duodenal neoplasms ; Neuroendocrine markers
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary A case of gangliocytic paraganglioma (GP) of the ampulla of Vater is reported and the literature reviewed, with special attention to immunohistochemical studies. The present case, which occurred in a 56-year-old woman, shows the typical histological admixture of epithelioid, ganglion and spindle cells. Immunohistochemistry reveals strong reactivity for synaptophysin, Leu-7, somatostatin, S-100 protein and vimentin. A few ganglion cells are reactive for neurofilaments. Chromogranin A, myelin basic protein, desmin and cytokeratin are absent. Immunohistochemical data from literature regarding the cytoskeletal composition of GPs are not unequivocal: cytokeratin and neurofilament positivity is reported by some authors and denied by others. More uniformity is reported concerning the peptides produced by GPs: somatostatin and pancreatic polypeptide are the most frequently found antigens, followed by serotonin. General neuroendocrine markers like neuron specific enolase and protein gene product 9.5 are always positive, whereas chromogranins are rarely found. S-100 protein is always positive in the spindle cell component. Our data are in keeping with those previously reported and add the diffuse positivity for the Leu-7 antigen and the positivity of ganglion cells for synaptophysin. The nature of the tumour is still a matter of debate and it is difficult to agree with either of the proposed hypotheses —hamartoma/choristoma versus true neoplasm. However the recent reports of the occasional malignant evolution of GPs may support their true neoplastic nature.
    Materialart: Digitale Medien
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  • 10
    Digitale Medien
    Digitale Medien
    Springer
    Journal of molecular medicine 67 (1989), S. 136-140 
    ISSN: 1432-1440
    Schlagwort(e): Immunocytochemistry ; Phospholipase A2 ; Pancreatic tumors ; Acinar-cell carcinoma
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Phospholipase A2 is an enzyme which is produced in acinar cells, and persists even in regressive states of chronic pancreatitis, when the production of other enzymes diminishes. We therefore tested this enzyme as a marker of acinar descent in various pancreatic tumors. This enzyme could be seen in 50% of the acinar-cell carcinomas, in 60% of solid and papillary pancreatic tumors, and in 50% of microglandular carcinomas. Ductal cancers and isletcell cancers were negative. In contrast to other markers of acinar matrix (amylase, antitrypsin), phospholipase A2 gave fewer false-positive or false-negative results.
    Materialart: Digitale Medien
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  • 11
    ISSN: 1432-0533
    Schlagwort(e): Medulloblastoma ; S-Antigen ; Rod-opsin ; Prognosis ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Biopsy specimens of 66 medulloblastomas were investigated by means of S-antigen and rod-opsin immunocytochemistry. The patients were operated between 1969 and 1988 and the medical records were retrospectively evaluated to correlate the immunocytochemical features of the tumors to the course of the disease. S-antigen- and rod-opsin-immunoreactive tumor cells were found in 19 out of 66 cases. Since in the normal non-neoplastic state immunoreactive S-antigen and rod-opsin are restricted to retinal photoreceptors and a class of pinealocytes derived from photoreceptor cells, the occurrence of these proteins in certain tumor cells of medulloblastomas suggests a differentiation of these cells along the photoreceptor cell lineage and allows the identification of a special subtype of medulloblastoma displaying photoreceptor-specific characteristics. This subtype appears to be closely related to retinoblastomas and pineal cell tumors. The incidence of this subtype corresponds to approximately 30% of all medulloblastomas. Correlation between the demonstration of immunoreactive S-antigen and rod-opsin and the course of the disease revealed a 10-year survival rate of 50.6% for patients with medulloblastomas displaying photoreceptor-specific characteristics and maximally 11% for patients suffering from medulloblastomas devoid of these markers. Although the statistical evaluation does not provide a significant result, the estimatedP-value of 0.085 indicates a distinct trend toward a better prognosis for patients suffering from medulloblastomas with photoreceptor-specific features. The validity of this trend needs to be proven in further studies with a greater number of patients.
    Materialart: Digitale Medien
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  • 12
    Digitale Medien
    Digitale Medien
    Springer
    Acta neuropathologica 77 (1989), S. 276-282 
    ISSN: 1432-0533
    Schlagwort(e): Cell kinetics ; Cell cycle ; Brain tumor ; Bromodeoxyuridine ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary A newly developed in vitro labeling method with bromodeoxyuridine (BrdU) identifies S phase cells in situ in freshly obtained surgical tissue of human brain tumors which is subsequently fixed and embedded in paraffin for BrdU immunovisualization. For the first time, the BrdU labeling index (LI) is successfully compared here with the LI obtained by immunostaining of frozen sections of the same tumors with monoclonal antibody Ki-67 which identifies all proliferating cells, i.e., the growth fraction. LIs were counted in at least five different areas with high density of labeled cells; at least 1,000 cells were counted. In 13 metastatic tumors, Ki-67 LI was 8.3%–62.6%, and BrdU LI was 5.1%–28.0%. In 18 gliomas, Ki-67 LI was 1.4%–19.3%, and BrdU LI was 0.2%–11.6%. In 7 meningiomas, Ki-67 LI was 0.3%–3.0%, and BrdU LI was 0%–2.0%. Statistical comparison of Ki-67 and BrdU LIs by linear regression analysis revealed a highly significant correlation: BrdU LI=0.99+0.34 Ki-67 LI (r=0.92,P〈0.001). A significant heterogeneity of proliferation patterns may occur within one sample from area to area, as well as between different samples of the same tumor, especially in gliomas; thus, some subjective influence on LIs by arbitrary sampling and selection could occur in quantitative evaluation of in situ cell kinetics of human brain tumors. This study indicates that our in vitro BrdU-labeling method allows the in situ identification of S phase cells in excellently preserved fixed tumor tissue which is well suited for further histological examination. This method compares favorably with Ki-67 labeling of frozen sections and might emerge as a powerful new tool for the routine study of cell proliferation in surgical specimens of human brain tumors.
    Materialart: Digitale Medien
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  • 13
    Digitale Medien
    Digitale Medien
    Springer
    Acta neuropathologica 77 (1989), S. 333-335 
    ISSN: 1432-0533
    Schlagwort(e): Canine distemper virus ; Astrocyte ; Immunocytochemistry ; Demyelination
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Acute canine distemper virus (CDV)-induced demyelinating lesions were examined with double-labelling immunocytochemistry simultaneously demonstrating CDV antigen and glial fibrillary acidid protein (GFAP) as marker for astrocytes. It was shown that 64% of all astrocytes within the demyelinating lesions were infected and that 95% of all infected cells counted in the lesions were astrocytes. These results suggest that the astrocyte ist the main target for CDV and that astroglial infection may play an important role in the mechanism of demyelination.
    Materialart: Digitale Medien
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  • 14
    ISSN: 1432-0533
    Schlagwort(e): Lewy bodies ; Pale bodies ; Neurofibrillary tangles ; Pick bodies ; Globose tangles ; Ubiquitin ; Antibodies ; Immunocytochemistry ; Alzheimer's disease ; Parkinson's disease ; Pick's disease ; Progressive supranuclear palsy
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Ubiquitin has previously been identified as a component of neuronal inclusions in neurodegenerative disorders. In this investigation, we examined tissue from cases of Alzheimer's disease (AD), Pick's discase, Parkinson's disease (PD), and progressive supranuclear palsy (PSP) to identify previously unrecognized ubiquitinated structures and to assess the evolution of neuronal inclusions. In AD, approximately 60% of neurofibrillary tangles (NFTs) that were stained with an anti-paired helical filaments (PHF) serum were identified by the ubiquitin antibodies. Extracellular NFTs were not labelled with anti-PHF but were unlabelled or weakly labelled with anti-ubiquitin antibodies. In Pick's disease, most Pick bodies were strongly labelled by the ubiquitin antibodies, and in addition some hippocampal CA1 neurones contained granular or strand-like ubiquitin-immunoreactive (IR) inclusions associated with more typical Pick bodies. Typical Lewy bodies in PD cases showed an unlabelled central core with an outer ring intensely labelled by ubiquitin antibodies. Pale bodies in pigmented substantia nigra neurones appeared as large well-defined, rounded structures without an identifiable core or peripheral zone. Some pale bodies were unlabelled by ubiquitin antibodies, but others showed labelling of variable intensity. Pale bodies which were labelled by ubiquitin antibodies tended also to be labelled by BF10, a monoclonal antibody against phosphorylated neurofilaments. We suggest that pale bodies in PD may represent stages in the formation of Lewy bodies. In addition, we observed numerous spindle-shaped ubiquitin-IR swellings of dendrites of pigmented substantia nigra neurones. In contrast to inclusions of AD and Pick's disease, the PHF-positive fibrillary neuronal inclusions of PSP were either unlabelled or only weakly labelled by ubiquitin antibodies. No ubiquitinated structures were seen in neurones from corresponding areas in aged controls. Identification of ubiquitinated proteins in neurodegenerative disorders may provide insights into molecular events associated with cell death.
    Materialart: Digitale Medien
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  • 15
    ISSN: 1432-0533
    Schlagwort(e): Alzheimer's disease ; Amyloid ; Immunocytochemistry ; Neuritic plaques ; Paired helical filaments
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Immunocytochemistry with monoclonal antibodies to the β-protein and to antigens associated with paired helical filaments (PHF) allows us to selectively stain two major components of neuritic (senile) plaques (NP): PHF and amyloid deposits. Using this method, the structure of NP in the brains of Alzheimer disease victims was compared to their structure in the brains of non-demented aged individuals selected for high numbers of NP. It is demonstrated that the dystrophic neurites participating in the plaque formation contain PHF only when cortical nerve cells in the same brain area form neurofibrillary tangles (NFT). People with many NP and many NFT were always demented, whereas people with many NP but few, if any NFT were not. It is speculated that there is individual susceptibility to the formation of PHF and that their appearance may represent a nonspecific response of the neuronal network to different kinds of injuries, like the deposition of amyloid in Alzheimer disease, or other pathogenic factors associated with various dementive neurodegenerative diseases. It is hypothesized that the deposition of brain amyloid in people resistant to neurofibrillary pathology may induced too little dysfunction for the development of dementia.
    Materialart: Digitale Medien
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  • 16
    Digitale Medien
    Digitale Medien
    Springer
    Acta neuropathologica 78 (1989), S. 497-503 
    ISSN: 1432-0533
    Schlagwort(e): Acquired immune deficiency syndrome (AIDS) ; Human immunodeficiency virus (HIV) ; Immunocytochemistry ; Vacuolar myelopathy ; Demyelination
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Vacuolar myelopathy (VM) is a frequent neurological complication of the acquired immune deficiency syndrome (AIDS). A suspected connection between VM and human immunodeficiency virus (HIV) has been based only on HIV isolation from affected spinal cord tissue. We report here an AIDS patient dying after 14 months of progressive dementia, including 3 months of spinal signs and symptoms. At autopsy, the brain revealed moderate diffuse damage of the white matter compatible with HIV-induced progressive diffuse leukoencephalopathy. The spinal cord showed VM mainly in the lateral and the posterior columns. Mono- and multinucleated macrophages were localized within intramyelinic and periaxonal vacuoles. Light and electron microscopic immunocytochemistry revealed the presence of HIV antigens restricted to mono- and multinucleated macrophages within the spongy lesions. Productive HIV infection is documented for the first time within VM lesions of this case. Therefore, VM should be included among HIV-induced lesions of the central nervous system. The intimate relation of infected macrophages to vacuolar myelinopathy could suggest secretion of a myelinotoxic factor by macrophages productively infected by HIV. Immune electron microscopy appears as promising tool to detect HIV in tissue even when the density of virus may be low.
    Materialart: Digitale Medien
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  • 17
    ISSN: 1432-0533
    Schlagwort(e): Pituitary adenoma ; Gangliocytoma ; Immunocytochemistry ; Pituitary hormone ; Neuropeptide
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Four tumors consisting of pituitary adenomatous cells (AD) intricated with ganglion cells (GC) were studied. Each case was associated with a different clinical syndrome: acromegaly, amenorrheagalactorrhea, Cushing's disease and isolated tumoral syndrome with no hormonal hypersecretion. (a) In the case with acromegaly, immunoreactive growth hormone (IR-GH) was present in 80% of AD. IR-vasoactive intestinal peptide (VIP) was found in 5%–10% of AD and in few GC. Rare GC and processes showed IR-GH-releasing hormone (GRH),-somatostatin (SRIH),-gonadotropin-relasing hormone and-adrenocorticotropin-releasing hormone. (b) In the case with amenorrhea-glactorrhea, IR-prolactin (PRL) was seen in 90% of AD. IR-PRL and-VIP were present in rare GC. (c) In the case with Cushing's disease, 60% of AD and very few GC contained IR-adrenocorticotropin (ACTH) and β-lipotropin. Rare GC processes contained IR-SRIH. (d) In the case without pituitary hormone hypersecretion, PRL was localized in rare AD and GC. Pituitary hormone and neuropeptides were nerver colocalized in the same cells. No case displayed IR-neurophysins or-thyroliberin. Pituitary hormones were localized by ultrastructural immunogold labeling. These findings show that: (i) in three cases, pituitary hormones (PRL and ACTH), and, in one case, VIP could be localized in both adenomatous and ganglion cells; (ii) the pituitary hormone-containing cells in the tumors could be related to the hypersecretory syndromes; (iii) intratumoral IR-VIP and-GRH might be involved in GH and PRL hypersecretion in the cases with acromegaly and amenorrhea-galactorrhea.
    Materialart: Digitale Medien
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  • 18
    Digitale Medien
    Digitale Medien
    Springer
    Acta neuropathologica 77 (1989), S. 441-444 
    ISSN: 1432-0533
    Schlagwort(e): Canine distemper virus ; Demyelination ; IgG index ; Encephalomyelitis ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary This is the first report of spontaneous canine distemper virus (CDV) infection in a dog associated with chronic progressive multiphasic neurological disease. Initial neurological deficits in the pelvic limbs progressed rapidly to paraplegia with almost complete remission after 9 weeks. Then another acute episode occurred with severe thoracic limb deficits and cerebellar dysfunction and progressive neurological deterioration over 3 months with rising serum neutralizing (SN) anti-CDV titers in the serum and cerebrospinal fluid (CSF). Three neuropathologically distinct lesions of spinal cystic necrosis, chronic demyelinating foci in the cerebellum and acute demyelinating encephalitis in the pons were identified. Persistent CDV antigen was demonstrated immunocytochemically only in acute lesions and atypically restricted to neurons. However, the immunological mechanism associated with the distinct remissions and exacerbations and CDV antigen clearance from chronic demyelinating lesions but persistence in acute lesions, despite a vigorous anti-CDV serologic response, was not defined.
    Materialart: Digitale Medien
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  • 19
    Digitale Medien
    Digitale Medien
    Springer
    Acta neuropathologica 79 (1989), S. 10-17 
    ISSN: 1432-0533
    Schlagwort(e): Locus ceruleus ; Pick bodies ; Lewy bodies ; Immunocytochemistry ; Electron microscopy
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary In classical Pick's disease with typical Pick bodies, inclusions resembling those present in the cerebral cortex are frequently found in the locus ceruleus. In three such cases Pick bodies were studied by light and electron microscopy and compared with Lewy bodies, inclusions more commonly found in this location. In contrast to the situation in the cerebral cortex, nerve cells with multiple Pick bodies were often found in the locus ceruleus, but in other respects definite light and electron microscopic differences between Pick bodies and Lewy bodies were present. Pick bodies were slightly basophilic and never had a central core or a peripheral halo. They were intensely argyrophilic. Differences in immunocytochemical reactions were especially marked with antibodies to tau and to paired helical filaments. Pick bodies displayed an intense reaction with these two antibodies, contrasting with that of Lewy bodies, which either lacked reactivity or reacted in a peripheral band. By electron microscopy the Pick bodies were composed of random filaments with smooth contour, whereas typical Lewy bodies had fuzzy deposits on filaments that radiated from a central core. Pick bodies in the locus ceruleus therefore maintained their immunocytochemical and electron microscopic characteristics and did not take on the character of Lewy bodies. Such differences point to a different pathogenesis and perhaps etiology of these two types of inclusions and attest to the marked difference clinically and pathologically between Pick's and Parkinson's diseases.
    Materialart: Digitale Medien
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  • 20
    Digitale Medien
    Digitale Medien
    Springer
    Acta neuropathologica 78 (1989), S. 337-347 
    ISSN: 1432-0533
    Schlagwort(e): Alzheimer's disease ; Amyloidosis ; β-protein ; Immunocytochemistry ; Paired helical filaments
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Immunocytochemical staining with monoclonal antibodies to the β-protein on tissue sections which have been pretreated with formic acid is not only a very specific but also a highly sensitive method for the detection of amyloid deposits in the brains of Alzheimer's disease victims. We report here a spectrum of morphological appearance of the brain amyloid deposits which are one of the main histopathological correlates of this disorder. Deposits of the β-protein are not only found in the well-known lesions [congophilic angiopathy and senile (neuritic) plaques] but are also seen under various morphological forms for which the word “plaques” does not appear an appropriate term: amyloid fibrils are found as large areas of diffuse infiltration of the neuropil, as ribbon-like infiltration in the subpial layer of the cerebral cortex, as granular deposits in the white matter, as diffuse deposits in the molecular layer of the cerebellum and the basal ganglia and as star-shaped deposits in the cerebellar Purkinje cell layer. The morphology of these deposits seems to depend on the cyto-and fibroarchitectonics of the brain region in which they are found, on the amount of amyloid deposited, and also on the type of staining technique used. It is only under specific circumstances that the deposition of amyloid in the neuropil is accompanied by the formation of paired helical filaments in nerve cell processes and their parent perikarya. In conclusion, our studies suggest that the extent of brain amyloidosis in Alzheimer's disease is much wider than so far appreciated.
    Materialart: Digitale Medien
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  • 21
    Digitale Medien
    Digitale Medien
    Springer
    Acta neuropathologica 78 (1989), S. 555-560 
    ISSN: 1432-0533
    Schlagwort(e): Sporadic motor neuron disease ; Lewy body-like hyaline inclusions ; Immunocytochemistry ; Electron microscopy
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Lewy body-like hyaline inclusions were immunocytochemically and electron microscopically investigated in a patient with sporadic motor neuron disease. The hyaline inclusions were chiefly observed within the perikarya of both normal-looking and chromatolytic anterior horn cells in the lumbar spinal cord, but some were detected in the axons and dendrites. Usually, a single inclusion was found in the perikaryon, but in rare cases two or more were observed. Immunocytochemically, these inclusions were intensely immunostained with anti-ubiquitin anti-body. Ultrastructurally, the hyaline inclusions were chiefly composed of randomly arranged linear structures associated with ribosome-like granules, varying from compactly arranged linear densities to more loosely packed ones. They contained scattered vesicles of various sizes and occasionally a focal accumulation of randomly arranged 10-nm neurofilaments or 13–25-nm filamentous structures.
    Materialart: Digitale Medien
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  • 22
    ISSN: 1432-0533
    Schlagwort(e): Cytomegalovirus (CMV) ; Acquired immunodeficiency syndrome (AIDS) ; Encephalitis ; DNA hybridization ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Brain tissues from 45 patients with AIDS and two brains with connatal cytomegalic inclusion body disease were investigated for a cytomegalovirus (CMV) etiology of encephalitic lesions. Nineteen brains showed evidence of CMV infection by histology, immunocytochemistry (ICC) using two different antibodies (mono- and polyclonal), and in situ hybridization (ISH). Fourteen cases with typical cytomegalic cells in conventional histology [eight with focally necrotizing encephalitis/ventriculitis including the two connatal infections and six with nodular encephalitis (NE)] revealed CMV with any method. In 5 of 15 AIDS cases of NE without cytomegalic cells, CMV infection was established by ISH, whereas ICC remained negative in these cases. Typical lesions of human immunodeficiency virus (HIV)-induced multifocal giant cell encephalitis (HIV encephalitis) in 13 brains were never labeled for CMV. In necrotizing encephalitis/ventriculitis, cell types which labeled for CMV, with and without cytomegalic change, comprised neurons, astrocytes, oligodendrocytes, ependyma, choroid plexus, endothelia, and cells in periand endoneurium, and in leptomeninges. Both ISH and ICC were able to detect widespread non-cytomegalic CMV-infected cells in normal parenchyma, well beyound the necrotizing lesions, in two AIDS cases. Labeling patterns of nuclei versus cytoplasms varied between the three methods for CMV detection. We conclude that in CNS tissues with cytomegalic cells, ICC and ISH are of comparable sensitivity; however, a diagnosis of CMV disease is possible in such cases by conventional histology. For an in situ diagnosis of CMV infection in NE without cytomegalic cells in AIDS, ISH is the method of choice. A selective vulnerability to CMV infection of any specific cell type of the human CNS is absent. With our detection methods, typical lesions of HIV encephalitis do not show local co-infection by CMV.
    Materialart: Digitale Medien
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  • 23
    Digitale Medien
    Digitale Medien
    Springer
    Acta neuropathologica 79 (1989), S. 340-346 
    ISSN: 1432-0533
    Schlagwort(e): Paraganglioma ; Melanin ; Orbit ; Immunocytochemistry ; Electron microscopy
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary A paraganglioma of the orbit in a 21-year-old woman is presented, containing oculo-cutaneous melanin in many tumor cells, occasionally adjacent to neurosecretory granules, and in macrophages. This tumor expands the list of neuroectodermal tumors with potential melaninization.
    Materialart: Digitale Medien
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  • 24
    Digitale Medien
    Digitale Medien
    Springer
    Anatomy and embryology 180 (1989), S. 1-15 
    ISSN: 1432-0568
    Schlagwort(e): Glutamate ; Taurine ; Glycine ; Immunocytochemistry ; Cerebellum ; Hippocampus ; Medulla spinalis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Amino acids are of crucial importance in brain function, not only as metabolic intermediates and building blocks of proteins, but also as mediators of interneuronal communication. This dual role of the amino acids distinguishes them from other neurotransmitter candidates, and implies that they are unlikely to be restricted to neurons using them as transmitters. This calls for a quantitative approach when attempts are made to analyse the distribution of transmitter amino acids by means of immunocytochemistry. The present review deals with recent methodological developments that have made it possible to utilize specific antisera to explore the cellular and subcellular distribution of neuroactive amino acids in a quantitative manner.
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  • 25
    Digitale Medien
    Digitale Medien
    Springer
    Anatomy and embryology 179 (1989), S. 221-226 
    ISSN: 1432-0568
    Schlagwort(e): GABA ; Immunocytochemistry ; Gastrointestinal tract ; Epithelium ; Enteroendocrine cells ; Rat
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Frozen sections of the corpus ventriculi, antrum pyloricum, duodenum, jejunum, ileum and colon from animals perfusion fixed with glutaraldehyde were treated with an antiserum specific for glutaraldehyde-fixed GABA and processed by the peroxidase antiperoxidase method. Semithin plastic sections from the antrum pyloricum were treated similarly. Stained cells appeared in the epithelium of all segments examined except the corpus ventriculi. The highest density of cells was observed along the major curvature of the antrum pyloricum. Here they were located in the bottom half of the gastric glands. Many of the cells showed a process extending towards the glandular lumen. No significant staining in the epithelium appeared when the antiserum was preincubated with glutaraldehyde-GABA complexes, nor when the anti-GABA serum was exchanged with anti-glycine or preimmune serum. The present findings and previous physiological data suggest that GABA may play a role in gut endocrine regulation.
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  • 26
    ISSN: 1432-0568
    Schlagwort(e): Syrian hamster ; Heart development ; endocardium ; Atrial natriuretic factor ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary The endocardium ultrastructure of 13 embryonic day old hamsters was examined, especially in relationship with the atrial myocytes. The endothelial morphology was described, including the junctional attachments and their relationships with subjacent atrial myocytes. Characteristic atrial myocytes organelles were identified: myofibrils, atrial granules, lipidic inclusions, and polysomes. Immunogold labeling demonstrated that atrial natriuretic factor (ANF)-containing granules were already present in the differentiating cardiomyocytes, even before the myofibrils were completely organized. At this stage of development, while the endothelium was a narrow barrier between the blood and the cardiomyocytes, it displayed fenestrations, but also epithelial discontinuities. In addition it also contains immunoreactive-ANF products. In light of the current knowledge about ANF processing it was proposed that the endocardium lining could be an obligated passageway for transport or activating proANF into ANF before its release into the blood stream. In addition the endocardial gaps could suggest that, until about 13 to 14 days of fetal development, heart atrial tissue could be more susceptible to the effects of pathogenetic compounds than in a later state of development.
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  • 27
    Digitale Medien
    Digitale Medien
    Springer
    Anatomy and embryology 179 (1989), S. 311-318 
    ISSN: 1432-0568
    Schlagwort(e): NCAM ; Thy-1 ; Immunocytochemistry ; Organs of special sense ; Mouse embryo
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary The distribution of the neural cell adhesion molecule (NCAM) and Thy-1 in the olfactory mucosa and olfactory bulb, the eye and the inner ear was examined with immunocytochemistry in mouse embryos from embryonic day 12 (E 12) to embryonic day 19 (E 19). In general, neurons are completely outlined with NCAM, whereas Thy-1 outlines only dendrites and axons. A variable cytoplasmic staining for Thy-1 is present in the perikarya. Neurons directly associated with special sense organs express NCAM and Thy-1 already from the earliest stage and throughout the period investigated, apart from the olfactory neurons in which Thy-1 disappears at E 19. The mitral cells in the olfactory bulb show Thy-1 but no NCAM reactivity. In the eye, lens fibers express Thy-1 and the pigmented layer expresses NCAM; neither of the two molecules can be detected at E 19. In the inner ear, hair cells express NCAM at E 19. Based on the distribution during the developmental period studied and on the cellular localisation of reaction products, it is suggested that the NCAM adhesion function could be of a more general nature by keeping appropriate cell membranes in close contact and thereby allowing more specific molecular interactions to take place. Thy-1, which is located on dendrites and axons, could be such a specific factor and function as recognition molecule in the developing nervous system.
    Materialart: Digitale Medien
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  • 28
    ISSN: 1432-0568
    Schlagwort(e): Atrial natriuretic peptide ; Pekin duck ; Salt gland ; Immunocytochemistry ; High performance liquid chromatography ; Radioimmunoassay
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary A novel peptide hormone, atrial natriuretic factor/cardiodilatin (ANP/CDD), was recently isolated and characterized from mammalian heart. Its presence has been demonstrated in several organs that contribute to water and sodium homeostasis, such as salivary glands. This study demonstrates the presence of ANP/CDD immunoreactivity in the salt gland of Pekin ducks by high performance liquid chromatography, radioimmunoassay and immunocytochemistry, using a specific antibody against atriopeptide I. A small number of distinct, ovoid or cuboid shaped ANP/CDD-immunoreactive cells were localized in the connective tissue surrounding and separating the central secretory tubules, whereas no immunostaining was observed in the peripheral tubules. Salt glands of ducks that were adapted to salt water revealed a significant hypertrophy of their secretory lobules. However, no differences were found between the number or localization of immunoreactive cells in the salt gland of salt water-acclimatized ducks and non-stimulated glands of ducks that were housed with ad libitum access to fresh water. Our results indicate that ANP/CDD may play a role in the regulation of sodium secretion in the salt gland of aquatic birds.
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  • 29
    ISSN: 1432-0568
    Schlagwort(e): Dogfish ; Peptides ; Immunocytochemistry ; Gastrointestinal ; Co-localisation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary The gastrointestinal tract of cartilaginous fishes, like that of higher vertebrates, is known to contain endocrine cells and nerves immunoreactive for a wide variety of peptides, some of which have been structurally characterised. Since we have found that substance P-, bombesin-and peptide histidine isoleucine-like immunoreactivities are similarly distributed in the endocrine cells of the dogfish pyloric stomach, we have tried to establish whether any of these peptides are co-localised. The cells were compared in thin serial sections with both ligh-and electron microscopical immunocytochemistry. Double immunolabelling was also used to show two immunoreactive peptides in the same tissue section. Further characterisation of the immunoreactivity was attempted by preabsorbing the antibodies with various peptides or synthetic fragments of peptide molecules. Immunoreactivity for all three peptides was frequently present in the same cells, whereas antibodies to other peptides such as gastrin and somatostatin marked different cells. Electron microscopy indicated that all the secretory granules in three morphologically different cell types reacted with antibodies to all three peptides. Dual localisation of unrelated peptides in endocrine cells or nerves is established in many cases, but triple localisation is as yet unusual. The immunoreaction for bombesin-like peptides is different in endocrine cells and nerves, indicating that dogfish bombesin may be present in two forms, in agreement with biochemical evidence.
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  • 30
    ISSN: 1432-1106
    Schlagwort(e): Kindling ; Inhibition ; GABA ; Immunocytochemistry ; Hippocampus
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary The kindling model of epilepsy, induced by tetanic stimulation of Schaffer collateral/commissural fibers, was studied in the rat hippocampus. Gamma-aminobutyric acid immunoreactivity was used to quantify the number of GABA-immunoreactive somata per mm2 in CA1 region, 28 days after the last generalized seizure. Comparison of the numbers obtained from kindled animals with those from controls, showed a significant decrease (18%) on the ipsilateral stimulated side but none on the contralateral side. In control rats injection of the GABA-transaminase inhibitor, amino oxyacetic acid (AOAA), led to a 46% increase in the number of cell somata immunoreactive for GABA. This probably results from an accumulation of GABA, reflecting GABA synthesis by glutamate decarboxylase (GAD) activity, in somata of interneurons that had initially a GABA content below the immunocytochemical detection threshold. In kindled rats, 31 days after the last seizure, the number of GABA-immunoreactive cells that could be observed after AOAA-treatment was significantly lower (35% ipsilateral and 25% contralateral) when compared to AOAA-treated controls. This suggests that in kindled animals a GAD dependent increase in GABA content did not take place in a subpopulation of interneurons. The observations for kindled rats are interpreted as a long-term decrease in GABA content and as an alteration in GABA turnover in a subpopulation of interneuron somata, the latter possibly due to a decrease in GAD activity. The long-term enhanced seizure sensitivity, characteristic for kindled animals, may be due to a decreased GABAergic inhibitory control of the neuronal circuitry in the CA1 region of the hippocampus.
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  • 31
    Digitale Medien
    Digitale Medien
    Springer
    Experimental brain research 78 (1989), S. 315-326 
    ISSN: 1432-1106
    Schlagwort(e): Efferents ; Labyrinth ; Immunocytochemistry ; Peptides
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary The efferent neurons of the gerbil vestibular system were investigated by retrograde tracing techniques and cytochemical staining for acetylcholinesterase (AChE), choline acetyltransferase (ChAT) and a number of peptides. The location, bilateral distribution, cell area and number of neurons in two identified groups of retrogradely labelled cells were described and quantified. The larger of the two groups was located dorsolateral to the facial nerve genu, ventral and medial to the vestibular nuclei. Unilateral tracer injection in the vestibular end organs labelled cells bilaterally in this and the smaller group, which was located immediately ventral to the genu. No cells were found that individually projected bilaterally to both labyrinths. After injections of horseradish peroxidase (HRP) in the utricle or saccule, significantly more cells were located on the contralateral side of the brainstem. The average (±SD) cross sectional area of labelled cell bodies associated with the otolith organs was 259.8 (±75.2) μm2. ChAT immuno-reactive and AChE positive cells were found in an area coextensive with the location of the dorsal efferent group. In double-labelling studies, cell bodies in the same group that had been retrogradely labelled with a utricular injection of HRP, were immunocytochemically stained for calcitonin generelated peptide and met-enkephalin. In contrast, the ventral group of efferents did not have cells that were cytochemically stained for either of the acetylcholine-related enzymes or either peptide. The significance of the existence of peptidergic vestibular efferent neurons is discussed.
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  • 32
    ISSN: 1432-1106
    Schlagwort(e): Nerve growth factor ; C6 glioma cells ; Chromaffin cells histofluorescence ; Electron microscopy ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary In order to maintain a chronic supply of growth factor for medulla cells in vitro, chromaffin cells from rat, African green monkeys and man were co-cultured with C6 glioma cells, which secrete growth factors that sustain sympathetic neurons in vitro. The response of chromaffin cells to coculture was compared to treatment of medullary cells with nerve growth factor (NGF) alone. Dispersed chromaffin cell preparations were obtained by a trypsin-collagenase procedure, and subjected to differential plating on collagen-coated surfaces. With both human and monkey tissue, non-chromaffin cells did attach to the culture plates and an enriched chromaffin cell population could be replated. Rat adrenal medulla cells survived very poorly in vitro and were not enriched in this procedure. Cultured human and monkey chromaffin cells survived as epithelial cells (50%) and showed neuritic outgrowth on 55 to 66% of the cells after eight days when treated with nerve growth factor (NGF). These cells showed strong catecholamine histofluorescence, tyrosine hydroxylase (TH) and dopamine beta hydroxylase (DBH) immunoreactivity. In contrast, only ten percent of adult rat chromaffin cells survived in culture, although NGF treatment rescued an additional 20% of the cells and induced neuritic outgrowth after one week in vitro. C6 glioma cells were treated with mitomycin C bromodeoxyuridine to inhibit mitosis and were plated with the various medulla cells in a one to one ratio. Both human and monkey chromaffin cells expressed extensive and enhanced neuritic arborization within eight days of co-culture, (64–82% respectively) and exhibited intimate contact with the glioma cells as seen at the ultrastructural level. Importantly, survival of adult rat adrenal medulla cells was enhanced to 50% or more with 40% of the cells extending neurites when co-cultured with glioma cells for seven days. Chromaffin cells from all three species reacted for TH, DBH and PNMT in co-culture and were histofluorescent. The majority of these cells were also immunoreactive for serotonin and enkephalin, while only 37% of chromaffin cells indicated the presence of NPY. These data indicate that adrenal medulla can be maintained in vitro as the neuronal phenotype when co-cultured with growth factor producing cells and that this strategy may be useful for in vivo transplantation studies.
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  • 33
    Digitale Medien
    Digitale Medien
    Springer
    Experimental brain research 78 (1989), S. 641-645 
    ISSN: 1432-1106
    Schlagwort(e): GABA ; Immunocytochemistry ; Pontine nuclei ; Baboon
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Using a postembedding immunogold procedure with an antiserum against glutaraldehyde-fixed GABA, we demonstrate GABA-like immunoreactivity in two classes of synaptic profiles in the pontine nuclei of baboon. One is an axon terminal in symmetrical synaptic contact with small or medium-sized GABA-immunonegative dendrites, the other is a pale, vesicle-containing profile resembling a dendrite or dendritic process which participates in serial synaptic arrangements. These synaptic arrangements, or triads, consist of a GABA-like immunoreactive, pale vesicle-containing profile being postsynaptic to a GABA-immunonegative axon terminal, and presynaptic to a small or medium-sized GABA-immunonegative dendrite. In at least some of these triads, the GABA-immunonegative axon terminals also contact the GABA-immunonegative dendrite directly.
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  • 34
    Digitale Medien
    Digitale Medien
    Springer
    Experimental brain research 78 (1989), S. 164-173 
    ISSN: 1432-1106
    Schlagwort(e): Glia ; GFAP ; Brainstem ; Spinal cord ; Immunocytochemistry ; Rat
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary The topographical mapping of glial fibrillary acidic protein (GFAP)-immunoreactivity was performed in coronal serial sections of the rat mesencephalon, rhombencephalon and spinal cord. Relative to a background of poor or moderate overall staining of the mesencephalon, the interpeduncular nucleus, substantia nigra and the periaqueductal grey matter were prominent by their intense GFAP-immunoreactivity. The pons and particularly the medulla contained more GFAP-labelled elements compared with the mesencephalon. The spinal trigeminal nucleus and Rolando substance were distinguished by their intense staining. Large fibre tracts were usually poor in immunoreactive GFAP. In a concluding discussion, findings relevant to the GFAP-mapping of the whole rat CNS are evaluated with regard to possible reasons underlying the observed differential distribution of GFAP-immunoreactivity.
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  • 35
    Digitale Medien
    Digitale Medien
    Springer
    Experimental brain research 76 (1989), S. 369-385 
    ISSN: 1432-1106
    Schlagwort(e): Cobalt experimental epilepsy ; Motor cortex ; GABA ; Immunocytochemistry ; Rat
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary The distribution of GABA-immunoreactive cell bodies and terminals was studied using an anti-GABA serum during the development of chronic focal epilepsy induced by cobalt deposits onto the motor cortex of the rat. Cell counts of GABA-positive neurons were carried out in the epileptogenic area and correlated with the electrophysiological activity of the cobalt focus. In normal control rats, we identified GABA-immunoreactive somata and processes in the motor agranular cortex; they were multipolar or bipolar but never pyramidal and were present in all layers, especially in layer II. GABA-immunoreactive terminals were widely scattered in the neuropil and surrounded the unlabelled cell bodies. In the cobalt-treated animals, changes in the GABAergic innervation were observed during the development of the epileptic focus: decreases in the GABA-positive cell density and in the number of GABA-positive terminals were present before the onset of epileptic discharges and became more marked during the period of maximal spiking activity; a progressive return to normal values of GABA-positive cell density (except in the deep layers) as well as the reappearance of GABA positive terminals were associated with the extinction of the epileptic syndrome. Our observations suggest that the impaired inhibitory neurotransmission mediated by GABA plays a role in the development of the cobalt-induced epilepsy; moreover the recovery of GABAergic function which occurs during the extinction of the epileptic syndrome might imply a capacity for axonal regeneration of the GABAergic neurons.
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  • 36
    Digitale Medien
    Digitale Medien
    Springer
    Experimental brain research 78 (1989), S. 479-488 
    ISSN: 1432-1106
    Schlagwort(e): Tachykinin ; Immunocytochemistry ; Thalamus ; Trigeminal ; Monkey
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Immunocytochemistry of fetal and adult monkey thalamus reveals a dense concentration of tachykinin immunoreactive fibers and terminals in the dorsolateral part of the VPM nucleus in which the contralateral side of the head, face and mouth is represented. The immunoreactive fibers enter the VPM nucleus from the thalamic fasciculus and electron microscopy reveals that they form large terminals resembling those of lemniscal axons and terminating in VPM on dendrites of relay neurons and on presynaptic dendrites of interneurons. Double labeling strategies involving immunostaining for tachykinins after retrograde labeling of brainstem neurons projecting to the VPM failed to reveal the origin of the fibers. The brainstem trigeminal nuclei, however, are regarded as the most likely sources of the VPM-projecting, tachykinin positive fibers.
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  • 37
    ISSN: 1432-0584
    Schlagwort(e): Immunocytochemistry ; Leukaemia ; Oncogenes
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary The expression of p21 ras proteins was investigated by immunocytochemistry in permanent cell lines and in fresh human leukaemic cells. While high and low levels of p21 ras could be detected in most of the cell lines, no significant p21 ras immunoreactivity was noted in cells of ten human acute and chronic leukaemias. Thus, notwithstanding its possible role in the initial transformation process in human leukaemias, p21 ras expression appears not to be an irrevocable requirement for the maintenance of the transformed state.
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  • 38
    ISSN: 1432-0428
    Schlagwort(e): Islet amyloid polypeptide ; Pancreatic islets ; B cells ; Ultrastructure ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Islet amyloid polypeptide is a novel 37 amino-acid-residues polypeptide which has been isolated from amyloid deposits in an insulinoma, and in human and cat islets of Langerhans. The molecule has 46% homology with the calcitonin gene-related peptide. Light microscopy examination of the pancreas shows that islet amyloid polypeptide immunoreactivity is restricted to the islet B cells. The present study utilized a rabbit antiserum against a synthetic peptide corresponding to positions 20–29 of islet amyloid polypeptide, a sequence without any amino-acid identity with calcitonin gene-related peptide. By applying the immunogold technique at the ultrastructural level, it was shown that both insulin and islet amyloid polypeptide immunoreactivity occurs in the central granular core of the human B cell secretory granules, while the A cells remain unlabelled. The demonstration that islet amyloid polypeptide is a granular protein of the B cells may indicate that it is released together with insulin. Further studies are necessary to evaluate the functional role of islet amyloid polypeptide.
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  • 39
    Digitale Medien
    Digitale Medien
    Springer
    Journal of neurology 236 (1989), S. 26-28 
    ISSN: 1432-1459
    Schlagwort(e): Acute viral meningoencephalitis ; Antigen-binding cells ; CSF ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary A method is presented that allows the immunocytochemical detection of varicella zoster virus antigen-binding cerebrospinal fluid cells in zoster ganglionitis. Antigenbinding cells were found only in patients suffering from this disease. The technique is sensitive, specific, inexpensive and relatively fast, and is potentially applicable to other inflammatory central nervous system diseases with immunoglobulin-containing cells (ICC) in CSF. The detection of antigen-binding CSF cells may represent a very early diagnostic test comparable with the early IgM antibodies of the systemic immune response.
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  • 40
    ISSN: 1432-1459
    Schlagwort(e): Nerve growth factor receptor ; Immunocytochemistry ; Monoclonal antibody ; Peripheral neuropathy ; Schwann cell
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary An analysis of nerve growth factor (NGF) receptor expression and density in human sural nerve biopsies was performed by immunocytochemistry with a murine monoclonal antibody against the human NGF receptor. Quantitative assessment of immunostaining density was made by histospectrophotometry on frozen sections. Although there was enhanced expression of NGF receptor within endoneurium in all patients with clinical neuropathies, expression was highest in nerves with axonal disease, consistent with the proposal that disruption of axon-Schwann cell interactions triggers the reexpression of the NGF receptor. These results with human nerves, together with previous studies with animal models, suggest that NGF and NGF receptor play important roles in the general response to neuronal injury.
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  • 41
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 258 (1989), S. 1-24 
    ISSN: 1432-0878
    Schlagwort(e): Serotonin ; Immunocytochemistry ; Insect nervous system ; Protocerebrum ; Suboesophageal ganglion ; Manduca sexta (Insecta)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Serotonin-immunoreactive neurons in the median protocerebrum and suboesophageal ganglion of the sphinx moth Manduca sexta were individually reconstructed. Serotonin immunoreactivity was detected in 19–20 bilaterally symmetrical pairs of interneurons in the midbrain and 10 pairs in the suboesophageal ganglion. These neurons were also immunoreactive with antisera against DOPA decarboxylase. All major neuropil regions except the protocerebral bridge are innervated by these neurons. In addition, efferent cells are serotonin-immunoreactive in the frontal ganglion (5 neurons) and the suboesophageal ganglion (2 pairs of neurons). The latter cells probably give rise to an extensive network of immunoreactive terminals on the surface of the suboesophageal ganglion and suboesophageal nerves. Most of the serotonin-immunoreactive neurons show a gradient in the intensity of immunoreactive staining, suggesting low levels of serotonin in cell bodies and dendritic arbors and highest concentrations in axonal terminals. Serotonin-immunoreactive cells often occur in pairs with similar morphological features. With one exception, all serotonin-immunoreactive neurons have bilateral projections with at least some arborizations in identical neuropil areas in both hemispheres. The morphology of several neurons suggests that they are part of neuronal feedback circuits. The similarity in the arborization patterns of serotonin-immunoreactive neurons raises the possibility that their outgrowing neurites experienced similar forces during embryonic development. The morphological similarities further suggest that serotonin-immunoreactive interneurons in the midbrain and suboesophageal ganglion share physiological characteristics.
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  • 42
    ISSN: 1432-0878
    Schlagwort(e): Met-enkephalin-Arg6-Phe7 ; Met-enkephalin-Arg6-Gly7-Leu8 ; Immunocytochemistry ; Neuropeptides ; Co-existence of peptides ; Neurosecretory cells, insects ; Blowfly, Calliphora vomitoria (Insecta)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Neuronal pathways immunoreactive to antisera against the extended-enkephalins, Met-enkephalin-Arg6-Phe7 (Met-7) and Met-enkephalin-Arg6-Gly7-Leu8 (Met-8), have been identified in the brain of the blowfly Calliphora vomitoria. Co-localisation with other enkephalins in certain neurons suggests that a precursor similar to preproenkephalin A exists in insects and that differential enzymatic processing occurs as in vertebrates. Co-localisations of the extended-enkephalin-like peptides with other vertebrate-type peptides, including cholecystokinin and pancreatic polypeptide, also occur. The enkephalinergic pathways are specific, comprising a few groups of highly characteristic neurons and areas of neuropil. Of special interest is the finding that parts of the antennal chemosensory and the optic lobe visual systems contain Met-8 immunoreactive neurons. Within the median neurosecretory cell groups, some of the giant neurons show immunoreactivity to Met-8 and others to both Met-8 and Met-7. Fibres from these cells project to the corpus cardiacum and also to the suboesophageal ganglion, where arborisations occur in the tritocerebral neuropil. Co-localisation studies of these cells have shown that at certain terminals, one particular type of peptide is the dominant neuroregulator, whilst at other terminals, within the same cell, a different co-synthesised peptide predominates. Several groups of lateral neurosecretory cells show clearly defined enkephalinergic pathways, most of which have connections with the central body. The complex patterns of immunoreactivity seen in terminals in the different parts of the central body, suggest an important role for the enkephalin-like peptides in the integration of multimodal sensory inputs. The physiological functions of the extended-enkephalin-like peptides in the brain of Calliphora is still unknown, but the anatomical evidence suggests they may have a role similar to that in mammals, where they are thought to control aspects of feeding behaviour.
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  • 43
    ISSN: 1432-0878
    Schlagwort(e): Gills ; Indoleamines ; Immunocytochemistry ; Autonomic innervation ; Salmo gairdneri R. ; Perca fluviatilis L. ; Micropterus dolomieui (Lacépède) ; Anguilla anguilla L. ; Ictalurus melas Rafinesque (Teleostei) ; Acipenser baeri L. (Chondrostei)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Antibodies against serotonin and 5-methoxytryptamine reveal indolaminergic neurons innervating the proximal part of the efferent arterial vasculature, the filament epithelia, the central venous sinus, and certain other serotonergic cells of the teleost gill filament. In the same area, acetylcholinesterase-positive and indoleaminergic neurons have already been described. We propose that these populations of neurons belong to a single neuronal type but express different agents. Our current results support this idea; in particular, they point to the presence of a single type of serotonin-containing nerve terminal, impinging on vascular smooth muscle. These results are in agreement with physiological data showing (i) the existence of non-cholinergic (atropine-resistant) vasoconstriction of the gill vasculature after nerve stimulation, and (ii) a potent vasoconstrictory action of infused serotonin. In addition, the above-mentioned serotonergic neurons have synaptic contacts with catecholaminergic nerve fibers, suggesting the existence of a modulatory relationship between the sympathetic and the cranial autonomic nerves supplying the teleost gill. Finally, these neurons show morphological relationships with a previously undescribed type of branchialserotonergic cell. The role of the parasympathetic nerve plexus of the teleost gill filament in the control of respiration and ionoregulation is discussed.
    Materialart: Digitale Medien
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  • 44
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 255 (1989), S. 657-662 
    ISSN: 1432-0878
    Schlagwort(e): Thyrotropin-releasing hormone (TRH) ; Development, ontogenetic ; Anterior hypothalamus ; Immunocytochemistry ; Domestic mallard
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Developmental changes of thyrotropin-releasing hormone (TRH)-immunoreactive structures in the brain of mallard embryos were studied by means of immunocytochemistry (PAP technique). The primary antibody was generated against synthetic TRH. Immunoreactive neurons were first detected in the hypothalamus of 14-day-old embryos. By day 20, increasing numbers of immunoreactive perikarya were observed in the paraventricular nucleus, anterior preoptic region and supraoptic region. Immunoreactive fiber projections were seen in the median eminence as early as embryonic day 20; they occurred also in some extrahypothalamic regions (lateral septum, accumbens nucleus). The number and staining intensity of the cell bodies increased up to hatching, and continued to increase during the first week after hatching.
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  • 45
    ISSN: 1432-0878
    Schlagwort(e): Development ; Intrapulmonary chemoreceptor ; Immunocytochemistry ; Morphometry ; Rabbit
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The distribution, frequency and size of neuroepithelial bodies (NEB) were studied in lungs of rabbits during different stages of development (27-day fetus, newborn, 6, 11, 21, 28 and 56 days postnatally). NEB were visualized by immunostaining with monoclonal antibody against serotonin. Detailed quantitiation of NEB was performed by use of camera lucida drawings of immunostained serial sections from the same anatomical region, i.e. the lower lobe of the left lung. The total number of NEB was counted and expressed per epithelial length of airway, surface area and volume. The size of NEB defined as surface area as well as the position of NEB in relation to the airway bifurcations was assessed in airways of different sizes. The overall number and size of NEB were found to increase during the immediate perinatal period followed by a sharp decline at 56 days of age. The number of NEB peaked at 6 days postnatally (mean 175.5 NEB/mm3 of airway epithelium) and declined significantly (3.0 NEB/mm3) at 56 days of postnatal age. The size of NEB reached its maximum at 11 days (mean surface area 659.54 μm2, with the largest NEB measuring 1839.98 μm2). By 56 days of age, NEB became significantly smaller (mean surface area 177.29 μm2) consisting of small clusters of cells situated deep within the airway epithelium. At all ages, about half of all NEB (mean 47.6%) were localized within the small peripheral airways with up to 63.9% located at airway bifurcations. These findings indicate that the “functional activity” of NEB may be confined predominantly to the perinatal period. The postulated functions of NEB include those of intrapulmonary hypoxia-sensitive chemoreceptors and/or endocrine-paracrine activity in the lung. Such function(s) may be important during adaptation to extrauterine life as well as for growth and development of the lung.
    Materialart: Digitale Medien
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  • 46
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 255 (1989), S. 523-527 
    ISSN: 1432-0878
    Schlagwort(e): Peptides ; Small intensely fluorescent (SIF) cell ; Neuropeptide coexistence ; Inferior mesenteric ganglion ; Immunocytochemistry ; Guinea pig
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Coexistence of peptides in the small intensely fluorescent cells was demonstrated by immunocytochemistry for met-enkephalin-Arg-Gly-Leu, vasoactive intestinal polypeptide, somatostatin, neuropeptide Y and dynorphin. In the extreme example, a single cell was immunoreactive to all 5 peptides examined. Four peptides coexisted in 8% and three peptides in 13% of SIF cells. In 10% of SIF cells no peptide immunoreactivity could be detected. The most prevalent peptide was met-enkephalin (in 46% of cells), then vasoactive intestinal polypeptide (45%), somatostatin (39%), neuropeptide Y (31%) and dynorphin (24%). Met-enkephalin and vasoactive intestinal polypeptide coexisted most commonly (25%).
    Materialart: Digitale Medien
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  • 47
    ISSN: 1432-0878
    Schlagwort(e): L1-antigen ; Cell adhesion molecule ; Developing vestibular neuroepithelium ; Immunocytochemistry ; Mouse (CBAxC57)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Doubts exist as to whether afferent nerve fibers exert a neurotrophic effect on the differentiation of sensory cells in the developing vestibular neuroepithelium. To determine whether innervation of hair cells precedes their differentiation, we have used the L1 adhesion molecule as a marker for axons. The detection of L1 on afferent axons in the otic vesicle of mouse embryos on gestation day 11 shows that nerve fibers penetrate the neuroepithelium before the sensory cells differentiate. L1-immunoreactivity of nerve endings also reveals the considerable fiber ramification on gestation days 14 and 15, i.e., corresponding to the first stages of sensory cell differentiation. The expression of L1 at successive stages of nerve fiber growth in the neuroepithelium, such as fasciculation and ramification, is not consistent with the previous role proposed for L1 as a fascicule-promoting factor and raises the possibility that other mechanisms are involved in L1 mediaded adhesion.
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  • 48
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 258 (1989), S. 101-109 
    ISSN: 1432-0878
    Schlagwort(e): Serotonin ; Immunocytochemistry ; Subesophageal ganglion ; Neurohemal organs ; Manduca sexta (Insecta)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Serotonin-immunoreactivity is mapped in wholemounts and slices of the suboesophageal ganglion (SOG) of larval Manduca sexta by means of immunocytochemistry. An extensive meshwork of serotonin-immunoreactive nerve fibres on some peripheral nerves of the SOG has been demonstrated. This meshwork appears to belong to a serotonergic neurohemal system, probably supplied by two pairs of bilateral serotonin-immunoreactive neurons with big cell bodies on the dorsal side near the midline in the mandibular neuromere. Intracellular recording and staining revealed their physiology and morphology. These neurons produce long lasting (50 msec) action potentials, which suggest that they are neurosecretory cells. Two pairs of bilateral serotonin-immunoreactive interneurons similar to those of other insects are stained in the labial and maxillar neuromeres, but not in the mandibular neuromere. Their ventrolaterally located cell bodies project through a ventral commissure into the contralateral hemiganglion and then cross back again through a dorsal commissure. The axons project into the contralateral circumoesophageal connective.
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  • 49
    ISSN: 1432-0878
    Schlagwort(e): Na+, K+-ATPase ; Immunocytochemistry ; Kidney ; Salivary glands ; Transport ; Rat (Sprague-Dawley)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary An antibody to the 96 kD α-subunit of the Na+, K+ -ATPase from Bufo marinus has been used in immunostaining rat kidney and salivary glands. Intense staining was observed on basolateral membranes of distal tubules of the kidney and striated ducts of the three major salivary glands. Less intense staining was seen on the basolateral membranes of parotid acinar cells, but no staining was seen on the acinar cells of submandibular or sublingual glands. These sites of staining have been shown, by other methods, to posses substantial Na+, K+ -ATPase, indicating that the antibody recognizes antigenic determinants of the sodium pump highly conserved in the course of evolution. In addition, staining with this antibody was observed at the apical region of cells of the proximal straight tubule and of the papillary collecting duct in the kidney. Absorption studies suggest that the apical antigenic determinants are the same or closely related to each other but are distinct from basolateral antigenic determinants.
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  • 50
    ISSN: 1432-0878
    Schlagwort(e): Subcommissural organ ; Secretory activity, neural control ; Transplantation ; Long-spacing collagen ; Immunocytochemistry ; Molecular markers (neuronal, glial) ; Electron microscopy ; Rat (Sprague-Dawley)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary There is increasing evidence that, in the rat, a serotonin-mediated neural input may have an inhibitory influence on the secretory activity of the subcommissural organ (SCO). In the present investigation the rat SCO was studied 7, 30 and 90 days after transplantation under the kidney capsule, an area devoid of local serotonin-containing nerves. The grafted tissue was examined by use of immunocytochemistry employing a series of primary antisera, lectin histochemistry and transmission electron microscopy. The grafted SCO survived transplantation and contained, in addition to secretory ependymal and hypendymal SCO-cells, also elements immunoreactive with antisera against glial fibrillary acidic protein or S-100 protein. In transplants, SCO-cells produced a material displaying the characteristic immunocytochemical and lectin-binding properties of SCO-cells observed under in-situ conditions. The ependymal cells lined 1–3 small cavities, which contained secretory material. A fully developed structural equivalent of Reissner's fiber was, however, never found. The immunocytochemical and ultrastructural study of the grafted SCO showed an absence of nerve fibers within the graft and suggested a state of enhanced secretory activity. A network of protruding basal lamina structures connected the secretory cells to the newly formed capillaries revascularizing the SCO. One week after transplantation, long-spacing collagen started to appear in expanded areas of such laminar networks and also in the perivascular space. It is suggested (i) that the formation of long-spacing forms of collagen is triggered by factors provided by the SCO-secretory cells, and (ii) that secretory material of the ependymal and hypendymal cells may reach the reticular extensions of the basal lamina. In contrast to the SCO in situ, the grafted SCO-cells showed a positive immunoreaction for neuron-specific enolase. They became surrounded by a S-100-immunoreactive glial sheath that separated them from other transplanted cell types and the adjacent kidney tissue of the host.
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  • 51
    ISSN: 1432-0878
    Schlagwort(e): Gut hormones ; Insulin ; Bombesin ; Immunocytochemistry ; Pancreatic polypeptide ; Cholecystokinin (CCK) ; Gastrin ; Schistocerca gregaria (Insecta)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The endocrine cells of the midgut epithelium of the desert locust are found dispersed among the digestive cells and are similar to those of the vertebrate gut. According to their reactivity to silver impregnation techniques and the ultrastructural features of the secretory granules (shape, electron-density, size, and structure) 10 types of endocrine cell have been identified, of which seven are located in the main segment of the midgut or in the enteric caeca, and the other three seem to be present only in the ampullae through which the Malpighian tubules drain into the gut. The endocrine cells have a slender cytoplasmic process that reaches the gut lumen, a feature that supports the receptosecretory nature postulated for this cellular type in insects as well as vertebrates. Antisera directed against mammalian gastrin, CCK, insulin, pancreatic polypeptide and bombesin reacted with some of the endocrine cells. This is the first time that insulin- and bombesin-like immunoreactive cells have been described in the midgut of an insect.
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  • 52
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 258 (1989), S. 119-124 
    ISSN: 1432-0878
    Schlagwort(e): Development, ontogenetic ; Immunocytochemistry ; Monoclonal antibodies ; Mucosa ; Lymphoid organs ; Domestic fowl
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The postnatal development of chicken mucosa-associated lymphoid tissues of the eyes, lungs, and intestines were investigated with monoclonal antibodies specific for either all leucocytes, B lymphocytes, mononuclear phagocytes, IgM, IgG, or IgA. Attention has been paid to the relation of lymphoid infiltrates with their surrounding mucosae, the segregation into B-cell and T-cell areas, development of germinal centers, and secretory immunoglobulins. Abudant secretory IgM and IgA was detected in the epithelium of the Harderian glands in the orbits, even though they lacked large leucocyte infiltrates with germinal centers. Lymphoid tissues in the mucosae of lungs and intestines developed separate B-cell and T-cell areas. The proventriculus, Meckel's diverticulum, and Peyer's patches generally contained germinal centers from 12 weeks of age on. Because chickens as young as 2 weeks old had germinal centers in bronchus-associated lymphoid tissue and cecal tonsils, these areas were probably highly stimulated by antigens. Isotype-specific monoclonal antibodies were used to detect IgM-, IgG-, and IgA-bearing follicular cells in the same germinal center.
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  • 53
    ISSN: 1432-0878
    Schlagwort(e): Somatotropes, growth hormone cells ; Immunocytochemistry ; Growth hormone (GH) ; Receptors, membrane ; Somatostatin (SRIF) ; Growth hormone-releasing hormone (GRH) ; Rat (Han: WIST)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The synthetic peptides somatostatin (SRIF) and growth hormone-releasing hormone (GRH) were coupled directly to colloidal gold of different particle sizes. Both conjugates were biologically active in displacing the corresponding radiolabeled hormones from high affinity binding sites in pituitary membranes. Release of growth hormone (GH) from cultured anterior pituitary cells was modulated by both conjugates alone or in combination. Ultrastructural studies were performed with cells incubated at 4° C (2 h) and 37° C (2 min-2 h) with one of the labeled peptides or their combination. Somatotropes were identified by immunostaining with anti-rGH followed by protein A-ferritin, thus obtaining a triple labeling. Both hormone conjugates were internalized in different vesicles in the beginning but accumulated during longer incubation times in the same compartment. The secretory vesicles and the nucleus were not labeled by any hormone conjugate. In contrast to SRIF-gold, the uptake of GRH-gold conjugate decreased with longer incubation times. This effect could be neutralized by simulatenous incubation of the somatotropes with both regulating hormones. Hence, whereas the binding and internalization of SRIF by somatotropes do not seem to be influenced by GRH, the corresponding processes for GRH are stimulated by the presence of SRIF.
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  • 54
    ISSN: 1432-0878
    Schlagwort(e): Insect AKH/RPCH ; Neurohormones ; Cam-HrTH-II ; Lom-AKH-I ; Immunocytochemistry ; Carausius morosus, Sarcophaga bullata (Insecta)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary A polyclonal antiserum was prepared against an N-terminal modified Cam-HrTH-II (Leu-Asn-Phe-...), one of the members of the large AKH/RPCH peptide family, first isolated from Carausius morosus. The localisation of this peptide was performed by means of immunocytochemical methods in the brain and corpora cardiaca-corpora allata complex of the stick insect, Carausius morosus and the grey fleshfly, Sarcophaga bullata. The distribution patterns of molecules reactive to the Cam-HrTH-II and the LomAKH-I antisera in both insect species were compared. In Carausius, both antisera reacted in the same cell bodies. In Sarcophaga, some neurons were stained by both, others only by one of the two antisera. By combining two different antisera, we demonstrated that there are no Lom-AKH-I-like molecules present in Carausius and that there must occur at least three different AKH-like molecules in the brain of Sarcophaga. One is similar to Cam-HrTH-II, the second to Lom-AKH-I and the third is an AKH/RPCH-like peptide, different from Lom-AKH-I and Cam-HrTH-II.
    Materialart: Digitale Medien
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  • 55
    ISSN: 1615-6102
    Schlagwort(e): Pyrenoid ; Ribulose 1,5-bisphosphate carboxylase/oxygenase ; Chlorella pyrenoidosa ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary The localization of ribulose 1,5-bisphosphate carboxylase/ oxygenase (RuBisCO) in cells ofC. pyrenoidosa grown at varying light intensities was determined by immunoelectron microscopy. Log phase cells grown at photon flux densities of 25 and 75 μEm−2s−1 (light-limiting) and 540 μEm−2s−1 (light-saturating) were fixed in 3% glutaraldehyde and embedded in Lowicryl K4M. Sections were labelled with antiserum to each subunit of RuBisCO followed by protein A-gold. At each light fluence rate, the pyrenoid was heavily labelled by each antibody whereas chloroplast stromal labelling was not above background levels. The apparent absence of stromal RuBisCO at each light level, and hence the lack of enzyme redistribution from pyrenoid to stroma following an increase in light fluence rate, suggests that pyrenoid RuBisCO is functional in vivo.
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  • 56
    ISSN: 1432-0878
    Schlagwort(e): Pituitary gland ; Neural lobe ; Intermediate lobe ; Intraventricular graft ; Immunocytochemistry ; Electron microscopy ; Rat (Wistar)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Neurointermediate lobes from adult or 10-dayold rats were implanted by a stereotaxic procedure into the third ventricle of adult male rats, in an area close to the paraventricular nucleus. They were examined, using immunocytochemical and ultrastructural techniques, at times ranging from 1 week to 8 months. All grafts were recovered in a healthy condition although some rejection of the tissue was detected at the 1and 2-week stages. In the neural lobe, clusters of pituicytes were scattered among the loose network of capillaries, most of which had a fenestrated endothelium. The intermediate lobe remained organized in compact avascular lobules. Axons similar to those projecting into the neurointermediate lobe in situ, but also axons of other types (e.g., somatostatinergic, enkephalinergic) penetrated the grafts. Synapses with melanotrophic cells in the intermediate lobe and neurohaemal contacts in the neural lobe were frequent from 2 1/2 months after transplantation. Immunocytochemical and ultrastructural characteristics indicated intense secretory stimulation of the melanotrophic cells in the early stages. All cells enclosed in a same glandular lobule reacted in a similar manner. In later stages, when re-innervation occurred, the cells recovered their initial characteristics. The overall effect of the re-innervation of the intermediate lobe grafted in this location is inhibitory, as in the lobe in situ.
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  • 57
    ISSN: 1432-0878
    Schlagwort(e): Spinal cord ; Dorsal root ganglia ; Skin ; Immunocytochemistry ; Neuropeptides ; Mutilated foot rat (Sprague-Dawley)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The mutilated foot rat is a mutant with autosomal recessive sensory neuropathy and frequent mutilation of the hindlimbs. Decreased numbers of dorsal root ganglion cells and diminished sensitivity to painful stimuli are characteristics of these animals. By use of immunocytochemistry, changes in the distributions of peptides involved in sensory and/or autonomic regulation, i.e. calcitonin generelated peptide (CGRP), tachykinins, enkephalin and neuropeptide Y in spinal cord, dorsal root ganglia and skin of these animals, were studied. In comparison with normal litter-mate controls, the dorsal horn of mutilated foot rats contained substantially fewer CGRP and tachykinin-immunoreactive fibres but more fibres immunoreactive for enkephalin. Many enkephalin-immunoreactive cell bodies were also found in the dorsal horn of the mutants, by contrast none were visible in control animals. Neuropeptide Y immunoreactivity was, however, unchanged in the spinal cord of the mutants. In the dorsal root ganglia of the mutants, the number of CGRPor tachykinin-immunoreactive cells and their proportion to total neuronal numbers were significantly less in comparison with normal controls. The diameter range of CGRP- and tachykinin-immunoreactive cells shifted from small (15–25 μm) to medium size (25–45 μm) as revealed by frequency distribution histograms. The skin from the affected foreand hindlimbs of the mutant rats, in keeping with fewer CGRP- and tachykinin-immunoreactive cells in the dorsal root ganglia, contained substantially less fibres immunoreactive for CGRP and tachykinins; a difference that was not seen in skin of unaffected areas (whiskers and snout). By contrast, neuropeptide Y-immunoreactive fibres showed a normal distribution around blood vessels and sweat glands of mutilated foot rats. The data suggest that diminished pain perception in the mutilated foot rat is related to loss of peptide-containing sensory neurones. Furthermore, the intraspinal increase of enkephalinergic neurones in the dorsal horn, concomitant with the decreased number of primary sensory neurones, may also play a contributory rôle in reducing pain thresholds.
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  • 58
    ISSN: 1432-0878
    Schlagwort(e): Olfactory-marker protein (OMP) ; Olfactory epithelium ; Immunocytochemistry ; Protein G-gold ; Freeze-substitution ; Lowicryl K11M embedding ; Rat (Sprague-Dawley)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary In this study electron-microscopic immunocytochemistry was used to localize olfactory marker protein in olfactory epithelia. Rat olfactory-epithelial samples were rapidly frozen, freeze-substituted with acetone, embedded at low temperatures with Lowicryl K11M and labelled on the sections with polyclonal antibodies raised against olfactory marker protein and with protein G conjugated to colloidal gold. Apart from the aforementioned use of acetone, substitution was carried out in the complete absence of chemical fixation, i.e., neither aldehydes nor OsO4 were used. This procedure resulted in localization concurrent with a good ultrastructural preservation. Olfactory-marker protein was present throughout the cytoplasmic compartments of dendrites and dendritic endings of olfactory-receptor cells, but it was not found in organelles such as mitochondria. Olfactory-marker protein was found only in dendriticendings of olfactory-receptor cells mature enough to have given rise to cilia, but these cilia displayed less labelling than dendrites and dendritic endings. Olfactory-marker protein was not found in apices and microvilli of neighboring olfactory-supporting cells.
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  • 59
    ISSN: 1432-0878
    Schlagwort(e): Thyroid gland ; Cathepsin B ; Lysosomes ; Immunocytochemistry ; Diurnal rhythm ; Rat (Wistar)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Immunocytochemical localization of cathepsin B and thyroxine (T4) in follicular cells of the rat thyroid gland and plasma concentrations of thyroid stimulating hormone (TSH) were examined at six evenly spaced times over 24 h. By light- and electron microscopy, immunodeposits for cathepsin B were localized in cytoplasmic granules of various sizes, whereas those for T4 were detected mainly in larger granules of the cells and in the colloid lumen. The size and location of cytoplasmic granules showing immunoreactivity for cathepsin B and T4 in the cells varied over 24 h, corresponding to a change in plasma TSH concentrations. These immunopositive large granules appeared in the apical cytoplasm at 12.00 h, when the level of TSH was highest. At 20.00 h when the level of TSH was lowest, T4-positive granules almost disappeared, and cathepsin B-positive small granules were abundantly seen in the basal region. From 00.00 h to 08.00 h, these positive granules changed in the same manner as those seen from 12.00 h to 20.00 h, associated with an increase in plasma TSH levels. These results suggest that newly formed colloid droplets migrate from the apical to the basal regions. Cathepsin B may play a role not only in the degradation of thyroglobulin but in the maturation of thyroid hormones during the migration of the granules.
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  • 60
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 256 (1989), S. 399-410 
    ISSN: 1432-0878
    Schlagwort(e): Dopamine ; Immunocytochemistry ; Brain, invertebrate ; Apis mellifera (Insecta)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The distribution of dopamine-like immunoreactive neurons is described for the brain of the bee, Apis mellifera L., following the application of a pre-embedding technique on Vibratome sections. Immunoreactive somata are grouped into seven clusters, mainly situated in the protocerebrum. Immunoreactive interneurons have been detected in the different neuropilar compartments, except for the optic lobe neuropils. Strong immunoreactivity is found in the upper division of the central body, in parts of the stalk and in the α-lobe layers of the mushroom bodies. A dense network of many immunoreactive fibres surrounds the mushroom bodies and the central body. It forms a number of interhemispheric commissures/chiasmata, projecting partly into the contralateral mushroom body and central body. The lateral protocerebral neuropil contains some large wide-field-neurons. The antennal-lobe glomeruli receive fine projections of multiglomerular dopamine-like immunoreactive interneurons.
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  • 61
    ISSN: 1432-0878
    Schlagwort(e): Pars distalis ; Prolactin/thyrotropic cells ; TRH ; Immunocytochemistry ; Morphometry ; Rana perezi (Anura)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The effects of synthetic thyrotropin-releasing hormone on pituitary prolactin and thyrotropic cells were investigated in adult male Rana perezi (formerly Rana ridibunda) frogs. Animals were given daily injections of synthetic thyrotropin-releasing hormone into the dorsal lymph sac. Prolactin and thyrotropic cells were identified by the colloidal-gold method, using anti-human prolactin and anti-human-β-thyrotropin hormone as primary antisera. The stereological parameters of the rough endoplasmic reticulum, Golgi complex, and secretory granules of prolactin and thyrotropic cells were evaluated by ultrastructural morphometry (point-counting method). Thyrotropin-releasing hormone caused cytological changes in both cell-types which were consistent with increased synthesis and release of both prolactin and thryrotropin. These changes were still significant after 48 h treatment in the case of thyrotropic cells, while in prolactin cells the thyrotropin-releasing hormone increased the number of secretory granules. After 6 days, the cells resembled essentially those used as controls. These results indicate that thyrotropin-releasing hormone stimulates the synthesis and release of prolactin and thyrotropin, and that the response of each cell type to this hypothalamic stimulus follows a different time-course.
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  • 62
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 257 (1989), S. 343-366 
    ISSN: 1432-0878
    Schlagwort(e): Central complex ; Golgi impregnation ; Neurotransmitters ; Protocerebrum, insect ; Immunocytochemistry ; Drosophila melanogaster (Insecta)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary On the basis of 1200 Golgi-impregnated brains the structure of the central complex of Drosophila melanogaster was analyzed at the cellular level. The four substructures of the central complex — the ellipsoid body, the fanshaped body, the noduli, and the protocerebral bridge — are composed of (a) columnar small-field elements linking different substructures or regions in the same substructure and (b) tangential large-field neurons forming strata perpendicular to the columns. At least some small-field neurons belong to isomorphic sets, which follow various regular projection patterns. Assuming that the blebs of a neuron are presynaptic and the spines are postsynaptic, the Golgi preparations indicate that small-field neurons projecting to the ventral bodies (accessory area) are the main output from the central complex and that its main input is through the large-field neurons. These in turn are presumed to receive input in various neuropils of the brain including the ventral bodies. Transmitters can be attributed immunocytochemically to some neuron types. For example, GABA is confined to the R1–R4 neurons of the ellipsoid body, whereas these cells are devoid of choline acetyltransferase-like immunore-activity. It is proposed that the central complex is an elaboration of the interhemispheric commissure serving the fast exchange of data between the two brain hemispheres in the control of behavioral activity.
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  • 63
    ISSN: 1432-0878
    Schlagwort(e): S-100 protein ; Immunocytochemistry ; Saccus vasculosus ; Pituitary gland, pars nervosa ; Mustelus manazo, Scyliorhinus torazame(Elasmobranchii)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary S-100 protein-immunoreactive cells were demonstrated by immunocytochemical procedures in the hypophysis and saccus vasculosus of two species of elasmobranchs (Mustelus manazo and Scyliorhinus torazame). In the saccus vasculosus of M. manazo, immunoreactivity was detectable exclusively in the fibrous portions interposed between the epithelial layer and the blood vessels. In the neurohypophysis, tanycytes and astrocytes of the median eminence were immunostained, but only a few labeled cells were found in the neurointermediate lobe. In S. torazame, the neurohypophysis displayed a similar distribution of immunoreactivity, but there were no labeled cells in the saccus vasculosus. In both species, none of the glandular cells of the hypophysis displayed immunoreactivity. Electron-microscopic examination showed that the immunostained cells in the saccus vasculosus correspond to astrocytes.
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  • 64
    ISSN: 1432-0878
    Schlagwort(e): Prolactin cells ; Growth hormone cells ; In situ hybridization ; Immunocytochemistry ; Cytogenesis ; Rat (Wistar-Imamichi)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Cytogenesis of growth hormone and prolactin cells in the rat pituitary gland was studied using in situ cDNA-mRNA hybridization and immunocytochemistry. Frozen or Paraplast sections of fetal and neonatal pituitaries were hybridized with 3H-cDNAs for rat prolactin or growth hormone, and were then processed for autoradiography. A number of growth hormone mRNA-positive cells were encountered throughout the anterior lobe on day 19 of gestation. Individual variaction in growth hormone gene expression was observed between fetuses at day 19 of gestation (6 out of 8 fetuses examined were positive for growth hormone mRNA). In contrast, growth hormone mRNA was detected in the all fetuses examined on day 20 or later. The autoradiographic signal (number of reduced silver grains) appeared to increase with later stages of development. Fetal growth hormone mRNA-positive cells were evenly scattered throughout the anterior lobe. Most of them were isolated, however, small clusters of several growth hormone cells were infrequently observed. Prolactin mRNApositive cells were found first on the 22nd day (the last day of gestation) in 3 of 6 fetuses examined, but were rarely observed on earlier gestational days. By postnatal day 8, prolactin mRNA-positive cells were numerous and the grain density over prolactin cells increased. Both growth hormone and prolactin cells were found as early as 18 days of gestation using immunocytochemistry, although the number of positive cells was very small at this stage. Immunoreactive growth hormone cells increased sharply in number during the next 24 h, while the number of prolactin cells remained scarce until birth. The results suggest that many growth hormone cells are still in an immature state at 20∶00 of day 18 and that many begin to synthesize growth hormone mRNA during next 14 h. On the other hand, no substantial prolactin gene expression appears to take place until after birth.
    Materialart: Digitale Medien
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  • 65
    ISSN: 1432-0878
    Schlagwort(e): Endocrine heart ; Estradiol ; Autoradiography ; Immunocytochemistry ; Co-localization ; CDD/ANP gene regulation ; Rat (Sprague-Dawley)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Atrial myoendocrine cells of rat were investigated regarding estradiol uptake. It was found that, in addition to their specific endocrine function of producing cardiac polypeptides of the cardiodilatin/atrial natriuretic peptide (CDD/ANP) family, these cells also specifically accumulate radiolabeled estradiol. This co-localization supports the view that steroid hormones play an important role in the regulation of the CDD/ANP gene.
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  • 66
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 256 (1989), S. 45-51 
    ISSN: 1432-0878
    Schlagwort(e): Testis ; Leydig cells ; Basement membrane ; Laminin ; Collagen ; Immunocytochemistry ; Rat (Wistar)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Rat testes were examined by conventional and immunolabeling transmission electron microscopy. Ultrastructurally identifiable continuous basement membranes were found around seminiferous tubules and the interstitial capillaries. Patches of basement membrane were, additionally, found on free surfaces of Leydig cells, between two Leydig cells, and in macrophage-Leydig cell contact sites. The ultrastructural findings were confirmed by immunocytochemical localization of laminin and collagen type IV in the same areas. A close association between the capillary basement membranes and the surfaces of perivascular Leydig cells was also observed. The possible basement membrane-mediated interactions of Leydig cells with other testicular structures, together with the novel bioactive products and regulators of Leydig cells, support the role of these cells as exceptionally complex regulatory centers of testicular functions.
    Materialart: Digitale Medien
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  • 67
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 255 (1989), S. 225-233 
    ISSN: 1432-0878
    Schlagwort(e): GABA ; Immunocytochemistry ; Visual system ; Optic lobes ; Periplaneta americana (Insecta)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Specific antisera against protein-conjugated γ-aminobutyric acid (GABA) were used in immunocytochemical staining procedures to study the distribution of the putative GABA-like immunoreactive neurons in the optic lobes of Periplaneta. GABA-like immunoreactive structures are evident in all three optic neuropil regions. Six different populations of GABAergic neurons, whose perikarya are grouped around the medulla, are found within the optic lobe. The number of these immunoreactive cells varies greatly and corresponds to the number of ommatidia of the eye. In the proximal part of the lamina, a coarse network of GABA-positive fibres is recognizable. These are the processes of large field tangential cells whose fibres pass through the distal surface of the medulla. A second fibre population of the lamina is made up of the processes of the centrifugal columnar neurons whose perikarya lie proximally to the medulla. The medulla contains 9 layers with GABAergic elements of variable immunoreactivity. Layers 1, 3, 5, 7 and 9 exhibit strong labelling, as a result of partial overlapping of the processes of centrifugal and centripetal columnar neurons, tangential fibres and/or lateral processes of perpendicular fibres and (possibly) processes of amacrines. A strong immunoreactivity is found in the proximal and distal layers of the lobula.
    Materialart: Digitale Medien
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  • 68
    ISSN: 1432-0878
    Schlagwort(e): Neuropeptides ; Immunocytochemistry ; Lumbricus terrestris (Annelida, Oligochaeta)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The localisation and distribution of 10 vertebrate-derived neuropeptides in the earthworm, Lumbricus terrestris, have been determined by an indirect immunofluorescence technique. The peptides are pancreatic polypeptide (PP), peptide tyrosine tyrosine (PYY), neuropeptide Y (NPY), glucagon (C-terminal), vasoactive intestinal polypeptide (VIP), peptide histidine isoleucine (PHI), gastrinreleasing peptide (GRP), calcitonin gene-related peptide (CGRP), neurotensin (NT), and met-enkephalin. For 6 of the peptides — PYY, NPY, PHI, glucagon, GRP and CGRP — this is the first demonstration of their presence in any annelid, and NT has not previously been described in an oligochaete. Cell bodies and nerve fibres immunoreactive to the 10 peptides occur throughout the CNS. In the PNS, epidermal sensory cells displayed immunoreactivities to PP and PYY, and PP-, PYY-, NPY-, PHI- and GRP-like immunoreactivities occurred in nerve fibres supplying the main body muscles. Nerve fibres immunoreactive to PP and PYY are also associated with the innervation of the gut (pharynx, oesophageal glands, and mid and posterior regions of the intestine). No endocrine cells immunoreactive for any of the antisera tested could be identified in the gut epithelium, suggesting that dual location of peptides in the brain and gut epithelium is a phenomenon that occurred at a later stage in evolution. No immunoreactive elements were detected in any of the organs and ducts of the reproductive and excretory systems.
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  • 69
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 257 (1989), S. 545-548 
    ISSN: 1432-0878
    Schlagwort(e): Brush cells ; Fatty acid-binding protein ; Immunocytochemistry ; Stomach ; Rat (Wistar)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary An immunocytochemical study by light- and electron microscopy using the antibody against rat hepatic fatty acid-binding protein (FABP) revealed the brush cells in the gastric epithelium of rats to be intensely immunoreactive. The immunoreactive cells were present in a group in the distal wall of the groove between forestomach and glandular stomach, as well as scattered singly in the surface and foveolar epithelia of the glandular stomach. Almost all immunoreactive brush cells had a thin basal process in contact with the basement membrane. No secretory granules with dense cores, similar to those of endocrine cells, were observed in the brush cells. The specific appearance of FABP-immunoreactivity in the brush cell indicates that this cell type is a distinct entity from other epithelial cells in the stomach and that FABP is a useful histochemical marker of the brush cells. FABP may be involved in the specific function(s) of this cell type related to fatty acid metabolism.
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  • 70
    ISSN: 1432-0878
    Schlagwort(e): Hypophyseal pars tuberalis ; (TSH), Thyrotropin ; Immunocytochemistry ; Photoperiod ; Phodopus sungorus
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Specific secretory cells in the hypophyseal pars tuberalis of Djungarian hamsters maintained under different photoperiods were investigated immunocytochemically by means of the colloidal gold technique using antibodies against rat thyrotropin (TSH). Secretory cells of animals kept under long photoperiods (LD16:8) showed positive staining of secretory granules (diameters 90–130 nm), whereas other intracellular structures were free of immunoreactivity. In animals kept under short photoperiods (LD8:16) secretory cells displayed increased numbers of secretory granules, but these organelles were devoid of immunoreactivity. In contrast, immunoreactivity of thyrotropes in the pars distalis did not differ between the two groups of animals investigated. The present results confirm earlier light-microscopical studies that in the pars tuberalis specific secretory cells show TSH-like immunoreactivity; however, they differ in their reactivity pattern from classical thyrotropes in the pars distalis.
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  • 71
    ISSN: 1432-0878
    Schlagwort(e): Neuropeptides ; Monoclonal antibodies ; Immunocytochemistry ; Endocrine system ; Nervous system ; Leptinotarsa decemlineata (Say)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Monoclonal antibodies were developed against peptidergic neurons in the nervous and endocrine tissues of the Colorado potato beetle by the immunization of mice with unpurified homogenates of these tissues. Methods were optimized to enhance chances for successful antibody production and selection, such as the pretreatment of the beetles, preparation of the immunogen, and screening hybridomas. Although only sub-microgram quantities of peptide antigen were used, many hybridomas generated antibodies recognizing peptidergic neurons in immunocytochemical procedures. A panel of 13 monoclonal antibodies anti-Colorado potato beetle (MACs) were harvested. All MACs stained different populations of peptidergic neurons, some of which had not been revealed by previously applied identification methods. Apart from the intrinsic glandular cells in the corpora cardiaca, immunoreactive neurosecretory neurons were revealed in medial and lateral groups in the protocerebrum and in the suboesophageal ganglion. These have axons terminating in the corpora cardiaca, and the neurosecretory granules can be revealed with the immunogold method. It is suggested that the immunoreactive substances represent neuropeptides or precursors of different kinds. Interneurons in other locations in the central and visceral nervous system have immunoreactive axonal projections that do not leave the ganglia. The set of MACs obtained is useful for neuroanatomical studies, for characterizing the secretory products, and for a further delineation of peptidergic communication channels in the insect body.
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  • 72
    Digitale Medien
    Digitale Medien
    Springer
    European archives of oto-rhino-laryngology and head & neck 246 (1989), S. 37-42 
    ISSN: 1434-4726
    Schlagwort(e): Cholesteatoma ; Cytokeratin ; Intermediate filaments ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary An indirect immunofluorescent method with monoclonal anti-cytokeratin antibodies was used to localize various cytokeratins in human middle ear cholesteatoma. The 50 K/58 K and 56.5 K/65–67 K paired cytokeratins are markers of skin type and were found in the specimens of human middle ear cholesteatomas studied. In contrast, the 40 K and 45 K cytokeratins (markers of simple epithelia), the 48 K cytokeratin (marker of hyperproliferative epidermal disease) and the 51K cytokeratin (marker of internal organ epithelia) were absent in human middle ear cholesteatoma. These findings indicate that the pattern of cytokeratins in human middle ear cholesteatoma is similar to that of skin but is different from those of simple epithelium, internal organ epithelia, and hyperproliferative epidermal disease. These findings also support the skin type epithelial origin of cholesteatoma and strongly favor the migration theory in the genesis of cholesteatomas.
    Materialart: Digitale Medien
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  • 73
    Digitale Medien
    Digitale Medien
    Springer
    European archives of oto-rhino-laryngology and head & neck 246 (1989), S. 222-226 
    ISSN: 1434-4726
    Schlagwort(e): Rhabdomyoma ; Immunocytochemistry ; Differential diagnosis
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Rhabdomyoma is a rare tumor of skeletal muscle origin which can occur in different localizations. The head and neck region is a site of predilection for the adult type. Its absolutely benign behavior makes correct diagnosis crucial. In this study, a case of an extracardiac rhabdomyoma of the larynx was found to recur 12 years after initial treatment. Following its surgical removal, histology showed typical patterns of an adult type of rhabdomyoma with focal cross-striations. Immunocytochemically, all applied muscular markers were positive. Electron microscopy revealed muscular filaments with cross-striations and glycogen granules. These findings showed that immunocytochemistry plays an important role in differential diagnosis and the need for possible further treatment.
    Materialart: Digitale Medien
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  • 74
    ISSN: 1432-0878
    Schlagwort(e): Gonadotropin-releasing hormone-associated peptide (GAP) ; Luteinizing hormone-releasing hormone (LHRH) ; Brain mapping ; LHRH prohormone ; Immunocytochemistry ; Rat (Wistar-R)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The gonadotropin-releasing hormone-associated peptide (GAP) of the LHRH precursor and the decapeptide LHRH were localized in the rat brain by immunocytochemistry in 12 to 18-day-old animals, by use of thick Vibratome sections and nickel intensification of the diaminobenzidinereaction product. Our results indicate that the GAP portion of the LHRH precursor is present in the same population of neurons that contain LHRH in the rat brain. An important difference observed was that the GAP antiserum, in contrast to LHRH antisera, stained several perikarya in the medial basal hypothalamus. GAP-immunoreactive perikarya were observed in the following regions: the olfactory bulb and tubercle, diagonal band of Broca, medial septum, medial preoptic and suprachiasmatic areas, anterior and lateral hypothalamus, and several regions of the hippocampus. In addition to the preoptico-terminal and the septopreoptico-infundibular pathways, we also observed GAPimmunopositive processes in several major tracts and areas of the brain, including the amygdala, stria terminalis, stria medullaris thalami, fasciculus retroflexus, stria longitudinalis medialis, periventricular plexus, periaqueductal gray of the mesencephalon and extra-cerebral regions, such as the nervus terminalis and its associated ganglion. These results confirm the specificity of previous immunocytochemical results obtained with antisera to LHRH. The presence of GAP immunoreactivity in nerve terminals of the rat brain indicates that GAP or a GAP-like peptide is located in the proper site to serve as a hypophysiotropic substance and/or as a neurotransmitter or neuromodulator.
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  • 75
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 256 (1989), S. 167-173 
    ISSN: 1432-0878
    Schlagwort(e): Cell proliferation ; Immunocytochemistry ; Lung ; Bronchioles ; Alveoli, lung ; Proliferating cell nuclear antigen ; Type II pneumocyte ; Clara cell ; Mouse (various strains)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Proliferating cell nuclear antigen is expressed in cells from late G1 through the S-phase of the cell cycle. Therefore, antibodies directed against this molecule should provide a probe for labeling immunocytochemically the nuclei of proliferating cells. Herein we demonstrate the feasibility and reliability of this technique by quantifying immunostained pulmonary nuclei. We applied polyclonal and monoclonal antisera to alveolar and bronchiolar pulmonary epithelial cells in various proliferative states in tissue-sections and in vitro. A/J mice had a slightly higher labeling index than C57BL/6J mice, and proliferation in both strains increased dramatically after butylated hydroxytoluene treatment produced compensatory hyperplasia of Type-II pneumocytes. Immunostaining in fetal and neonatal lung samples from mice was higher than in adults. Spontaneous lung adenomas had a higher labeling index than the surrounding normal lung tissue. In addition, new data contained herein demonstrate a strain difference in proliferation of bronchiolar epithelial cells, and quantify the extent to which BHT-induced lung damage increases these proliferative rates. This mammalian nuclear antigen did not cross-react with antiserum to a functionally related bacterial protein, the beta subunit of E. coli DNA polymerase-III holoenzyme.
    Materialart: Digitale Medien
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  • 76
    ISSN: 1432-0878
    Schlagwort(e): Insulin-related peptide ; Immunocytochemistry ; Immunochemical characterization ; Pancreas ; Ambystoma mexicanum (Urodela)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The pancreas of the axolotl, Ambystoma mexicanum, was investigated by immunocytochemical methods for the presence of immunoreactivity to a number of antisera raised against mammalian insulins. All anti-insulin antisera tested revealed substantial amounts of reaction products confined solely to the aldehyde-fuchsinophilic B cells of the endocrine pancreas. The reactive cell population was detected by use of one polyclonal antiserum against bovine insulin and eight different monoclonal antibodies against insulins from various mammalian species. Six of these antibody clones have known specificity to sub-regions of the insulin molecule. Additionally, fractions of an ethanol-HCl extract of pancreatic tissue from Ambystoma was studied in both conventional dot-blot tests by means of the same panel of antibodies and a two-site sandwich time-resolved immunofluorometric assay for human insulin involving two of the monoclonal antibodies. These experiments support the immunocytochemical observations by demonstrating the existence of an insulin-related peptide with a great deal of structural resemblance to mammalian insulins and displaying antigenic determinants in common at least with the amino acid residues A8–10 and B26–30. In conclusion, we interpret the findings as indicating that the immunocytochemically revealed tissue bound antigen in the Ambystoma pancreatic B-cells may be a peptide related to human insulin.
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  • 77
    ISSN: 1432-0878
    Schlagwort(e): Immunocytochemistry ; Prolactin cells ; Gonadotropic cells ; ACTH cells ; Folliculo-stellate cells ; Rana pipiens (Anura)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The colloidal gold immunocytochemical technique was used to determine the ultrastructural features of the glandular cells in the pituitaries of male frogs, Rana pipiens, both in vivo and after superfusion in vitro. Specific reactions to antisera against bullfrog gonadotropins, human prolactin, and synthetic 1–39 corticotropin allowed identification of the 3 corresponding types of glandular cells. No immunoreaction was obtained with antisera against human or ovine-growth hormone, human β-thyrotropin hormone, and bovine S-100 protein. General morphological features of these immunocytochemically identified glandular cells were similar to those of equivalent cells previously described in other amphibian species. Non-glandular folliculo-stellate cells were distinctive. In freshly removed pituitaries, these folliculo-stellate cells contained lysosome-like structures, but did not show phagocytic vacuoles in the cytoplasm; they contained many mitochondria, and the Golgi complex and endoplasmic reticulum were relatively undeveloped. After 4 or 18 h of superfusion, some immunoreactive gonadotropic, prolactin, and corticotropic cells showed degeneration and destruction. In the same gland, folliculo-stellate cells retained a viable appearance, but showed phagocytic vacuoles containing secretory granule-like structures which were immunoreactive to gonadotropic, prolactin, and corticotropic antibodies. Some folliculo-stellate cells showed phagocytic vacuoles containing complete glandular cells. These results suggest that superfusion causes a destruction of some of the glandular cells, and that folliculo-stellate cells act as phagocytes when cellular debris or moribund cells are present in the intercellular space in the pituitary parenchyma.
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  • 78
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 257 (1989), S. 313-323 
    ISSN: 1432-0878
    Schlagwort(e): Serotonin (5HT) ; Immunocytochemistry ; 5,6-Dihydroxytryptamine ; Central nervous system ; Helix pomatia (Mollusca)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The distribution of serotonin (5HT)-containing neurons in the central nervous system of the snail Helix pomatia has been determined in whole-mount preparations by use of immunocytochemical and in vivo 5,6-dihydroxy-tryptamine labelling. 5HT-immunoreactive neuronal somata occur in all but the buccal and pleural ganglia. Immunoreactive fibres are present throughout the central nervous system. The 5HT-immunoreactive neuronal somata characteristically appear in groups, located mainly in the cerebral, pedal, visceral and right parietal ganglia. The majority of 5HT-immunoreactive neurons is located in the pedal ganglia. Additionally a dense network of 5HT-immunoreactive varicose fibres is found in the neural sheath of the central nervous system including all the nerves and ganglia. The number and distribution of 5HT-immunoreactive neurons correlates with that demonstrated by 5,6-dihydroxytryptamine labelling method.
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  • 79
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 255 (1989), S. 215-224 
    ISSN: 1432-0878
    Schlagwort(e): Transglutaminase ; Immunocytochemistry ; Induction ; Tissue compartments ; Tissue integrity ; Human
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Tissue transglutaminase is an intracellular enzyme without established physiological function. Biochemically it can be detected in all organs, but no systematic in situ localization has been carried out so far. Here we report the immunohistochemical localization of transglutaminase in human tissues using an affinity purified, monospecific anti-human transglutaminase antibody. It is shown that the widespread organ distribution of the enzyme is the consequence of its occurrence in ubiquitous cell types such as endothelium and smooth muscle cells. Some organ-specific cell types express the enzyme constitutively (mesangial cells, renomedullary interstitial cells, thymic subcapsular epithelium, colonic pericryptal fibroblasts), while in others it seems to be induced either by external stimuli (epithelium of the female breast) or as part of their differentiation/maturation program (developing nephrons, enterocytes of the small intestine). The presence of tissue transglutaminase can be demonstrated in derivatives of all germ layers and in the trophoblast. The functional implications of these findings are presently unknown; however, based on its distribution the role of this enzyme in compartmentation and preservation of tissue integrity against stress may be suggested.
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  • 80
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 256 (1989), S. 559-565 
    ISSN: 1432-0878
    Schlagwort(e): Retrograde tracing ; Immunocytochemistry ; Vascular innervation ; Rat
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The origin of nerve fibers to the superficial temporal artery of the rat was studied by retrograde tracing with the fluorescent dye True Blue (TB). Application of TB to the rat superficial temporal artery labeled perikarya in the superior cervical ganglion, the otic ganglion, the sphenopalatine ganglion, the jugular-nodose ganglionic complex, and the trigeminal ganglion. The labeled perikarya were located in ipsilateral ganglia; a few neuronal somata were, in addition, seen in contralateral ganglia. Judging from the number of labeled nerve cell bodies the majority of fibers contributing to the perivascular innervation originate from the superior cervical, sphenopalatine and trigeminal ganglia. A moderate labeling was seen in the otic ganglion, whereas only few perikarya were labeled in the jugular-nodose ganglionic complex. Furthermore, TB-labeled perikarya were examined for the presence of neuropeptides. In the superior cervical ganglion, all TB-labeled nerve cell bodies contained neuropeptide Y. In the sphenopalatine and otic ganglia, the majority of the labeled perikarya were endowed with vasoactive intestinal polypeptide. In the trigeminal ganglion, the majority of the TB-labeled nerve cell bodies displayed calcitonin gene-related peptide, while a small population of the TB-labeled neuronal elements contained, in addition, substance P. In conclusion, these findings indicate that the majority of peptide-containing nerve fibers to the superficial temporal artery originate in ipsilateral cranial ganglia; a few fibers, however, may originate in contralateral ganglia.
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  • 81
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 257 (1989), S. 69-75 
    ISSN: 1432-0878
    Schlagwort(e): Inner ear ; Cytoskeletal proteins ; Immunocytochemistry ; Cell motility ; Actin ; Guinea pig
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary Isolated outer hair cells of the guinea pig lacking a cuticular plate and its associated infracuticular network retain the ability to shorten longitudinally and become thinner. Membrane ghosts lacking cytoplasm retain the cylindrical shape of the hair-cell, and although they do not shorten, they retain the ability to constrict and become thinner. These data suggest that cytoplasmic components are associated with outer hair-cell longitudinal shortening and that the lateral wall is responsible for maintaing cell shape and for constriction. Actin, a protein associated with the cytoskeleton and cell motility, is thought to be involved in outer hair-cell motility. To study its role, actin was localized in isolated outer hair cells by use of phalloidin labeled with fluorescein and antibodies against actin coupled to colloidal gold. In permeabilized guinea-pig hair cells stained with phalloidin, actin filaments are found along the lateral wall. In frozen-fixed hair cells actin filaments are distributed uniformly throughout the cytoplasm. Electron-microscopic studies show that antibodies label actin throughout the outer hair-cell body. Thus cytoplasmic actin filaments may provide the structural basis for the contraction-like events.
    Materialart: Digitale Medien
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  • 82
    ISSN: 1432-0878
    Schlagwort(e): Antigen localization ; FMRFamide-like immunoreactivity ; Immunocytochemistry ; Invertebrate ganglia ; Small cardioactive peptide-like immunoreactivity ; Hirudo medicinalis (Annelida)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The distributions of small cardioactive peptide (SCP)- and FMRFamide-like immunoreactivities in the central nervous system of the medicinal leech Hirudo medicinalis were studied. A subset of neurons in the segmental ganglia and brains was immunoreactive to an antibody directed against SCPB. Immunoreactive cell bodies were regionally distributed throughout the nerve cord, and occurred both as bilaterally paired and unpaired neurons. The majority of the unpaired cells displayed a tendency to alternate from side to side in adjacent ganglia. A small number of neurons were immunoreactive only in a minority of nerve cords investigated. Intracellular injections of Lucifer yellow dye and subsequent processing for immunocytochemistry revealed SCP-like immunoreactivity in heart modulatory neurons but not in heart motor neurons. FMRFamide-like immunoreactivity was also detected in cell bodies throughout the central nervous system. A subset of neurons contained both SCP- and FMRFamide-like immunoreactivities; others stained for only one or the other antigen. These data suggest that an antigen distinct from FMRFamide is responsible for at least part of the SCP-like immunoreactivity. This antigen likely bears some homology to the carboxyl terminal of SCPA and SCPB.
    Materialart: Digitale Medien
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  • 83
    ISSN: 1432-0878
    Schlagwort(e): Subcommissural organ ; Glial markers ; Immunocytochemistry ; GABA uptake ; Comparative analysis ; Mammals (rat, cat, mouse, rabbit)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The subcommissural organ (SCO), classified as one of the circumventricular organs, is composed mainly of modified ependymal cells, attributable to a glial lineage. Nevertheless, in the rat, these cells do not possess glial markers such as glial fibrillary acidic protein (GFAP), protein S100, or the enzyme glutamine synthetase (GS). They receive a synaptic 5-HT input and show pharmacological properties for uptake of GABA resembling the uptake mechanism of neurons. In this study, we examine the phenotype of several mammalian SCO (cat, mouse, rabbit) and compare them with the corresponding features of the rat SCO. In all these species, the SCO ependymocytes possess vimentin as an intermediate filament, but never express GFAP or neurofilament proteins. They do not contain GS as do glial cells involved in GABA metabolism, and when they contain protein S100 (rabbit, mouse), its rate is low in comparison to classical glial or ependymal cells. Thus, these ependymocytes display characteristics that differentiate them from other types of glial cells (astrocytes, epithelial ependymocytes and tanycytes). Striking interspecies differences in the capacity of SCO-ependymocytes for uptake of GABA might be related to their innervation and suggest a species-dependent plasticity in their function.
    Materialart: Digitale Medien
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  • 84
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 256 (1989), S. 153-158 
    ISSN: 1432-0878
    Schlagwort(e): Acetylcholine ; Acetylcholinesterase ; Cholin-acetyltransferase ; Immunocytochemistry ; Visual system ; Calliphora erythrocephala (Insecta)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The distribution of putative cholinergic neurons in the lamina of the blowfly Calliphora erythrocephala was studied by immunocytochemical and histochemical methods. Three different antibodies directed against the AChsynthesizing enzyme, choline acetyltransferase (ChAT), revealed a cholinergic population of fibres running parallel to the laminar cartridges, which have branch-like structures at the distal lamina border. Cell bodies in the chiasma next to the lamina border were also labelled by the anti-ChAT antibodies. Monopolar cell bodies in the nuclear layer were faintly labelled. The distribution of the acetylcholine hydrolyzing enzyme, acetylcholine esterase (AChE), was revealed by histochemical staining and was similar to the ChAT immunocytochemistry. The arrangement of ChAT positive fibres in transverse and longitudinal sections and the distribution of AChE stained fibres indicate that the amacrine cells of the lamina are cholinergic cells.
    Materialart: Digitale Medien
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  • 85
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 256 (1989), S. 159-166 
    ISSN: 1432-0878
    Schlagwort(e): Pancreas, endocrine ; Cysteamine ; Somatostatin ; Insulin secretion ; Glucose ; Immunocytochemistry ; Mouse (NMRI)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary To evaluate the previously reported depletion of pancreatic somatostatin by cysteamine (β-mercaptoethylamine), mice were injected subcutaneously with the drug at 300 mg/kg. Immunocytochemical analysis performed on sections from tissue taken at 4 h after the injection revealed an elimination of somatostatin-14-like immunoreactivity without alterations in the somatostatin-28(1 – 12)-like immunoreactivity. In sections from tissues taken at 24 h after injection, no differences between cysteamine-injected animals and controls were observed. Immunochemical analysis of somatostatin-14-like immunoreactivity in pancreatic extracts showed a significant reduction of the concentration (P〈 0.001). In contrast, no change in the insulin concentration was observed. Functionally, cysteamine lowered the plasma glucose levels at l h after injection; this effect persisted for 6 h. Plasma insulin levels were likewise reduced transiently by cysteamine. Concomitant administration of somatostatin did not influence these effects of cysteamine. The plasma glucose-lowering effect of cysteamine was seen also in alloxan-diabetic mice. We conclude that cysteamine alters the immunoreactive characteristics of pancreatic somatostatin without affecting the immunoreactivity of insulin, and that cysteamine transiently reduces plasma glucose and insulin levels
    Materialart: Digitale Medien
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  • 86
    ISSN: 1432-0878
    Schlagwort(e): FMRFamide ; Cholecystokinin ; Immunocytochemistry ; Insect visual system ; Neural development ; Calliphora erythrocephala (Insecta)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The adult optic lobes of the blowfly Calliphora erythrocephala were found to be innervated by more than 2000 neurons immunoreactive to antisera raised against the neuropeptides FMRFamide, its fragment RFamide, and gastrin/cholecystokinin (CCK). All of the CCK-like immunoreactive (CCK-IR) neurons also reacted with antisera to RFamide, FMRFamide and pancreatic polypeptide. A few RFamide/FMRFamide-like immunoreactive (RF-IR) neurons did not react with CCK antisera; they reacted instead with antisera to Leu-enkephalin and Met-enkephalin-Arg6-Phe7. The RF-IR neurons are, thus, heterogeneous with respect to their contents of immunoreactive peptides. Two of the RF-IR neuron types innervating the adult optic lobes could be traced in their entirety only after following their postembryonic development, because of the complexity of the trajectories of the immunoreactive neuronal process in the adult insect. The majority of the cell bodies of the RF-IR and CCK-IR neurons lie within the optic lobes and are derived from imaginal neuroblasts of the inner and outer optic anlagen. Six of the peptidergic neurons are, however, metamorphosing larval neurons with their cell bodies in the central part of the protocerebrum. The full extent of immunoreactivitiy is not attained in some of the neurons until the late pupal or early adult stage. The larval optic center was also found to be innervated by neurons immuno-reactive with both RFamide and CCK antisera. The cell bodies of these RF-IR/CCK-IR neurons are located near the developing lamina (one on each side). In the 24 h pupa, the cell bodies of these neurons are still immunoreactive, but thereafter they cannot be immunolabeled apparently due to cell death or a change in transmitter phenotype.
    Materialart: Digitale Medien
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  • 87
    Digitale Medien
    Digitale Medien
    Springer
    Cell & tissue research 255 (1989), S. 645-650 
    ISSN: 1432-0878
    Schlagwort(e): Pituitary, pars distalis ; Differentiation ; Organ culture ; Immunocytochemistry ; Rat (Sprague-Dawley, CFY)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary An immunohistochemical study was performed to determine the capacity of early fetal pituitaries to differentiate into specific hormone-synthesizing tissue in the absence of any influence from the central nervous system. Rathke's pouches from rats were removed from their juxtadiencephalic position on day 11 and 12 of gestation and maintained for 2–7 days in a chemically defined culture medium (M 199) without antibiotics and serum supplementation. The immunocytochemical observations provided evidence for the differentiation of ACTH-, TSH-gb-, LH-gb-, FSH-gb-, GH- and PRL-synthesizing cells in the isolated organ cultured from 11 to 12-day-old pituitary primordia. The appearance of specific hormone-synthesizing cells in vitro displayed a delay of 1.5–2 days compared to the day of appearance in vivo, however, the sequential order of developmental events occurred as observed in vivo. The present results suggest that endocrine or neuroendocrine signals are not required for the expression of specific secretory functions of fetal pituitaries, at least at an age of 11–12 days.
    Materialart: Digitale Medien
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  • 88
    ISSN: 1432-0878
    Schlagwort(e): Nucleus intercollicularis ; α2-Adrenergic receptors ; Estrogen receptors ; Quantitative autoradiography ; Immunocytochemistry ; Japanese quail, Coturnix coturnix japonica (Aves, Phasianiformes)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Summary The nucleus intercollicularis is an important site in the control of vocalization in birds. In oscines, a subregion of the nucleus intercollicularis called the dorso-medial intercollicular nucleus appears to play a key role in this process because it receives the majority of the projections from the nucleus robustus archistriatalis and sends most of the projections to the motor nucleus of the hypoglossal nerve. In this paper, we present neurochemical studies of the nucleus intercollicularis in the Japanese quail which suggest the presence of heterogeneity within this structure. One rostral band contains high densities of cholinergic muscarinic receptors identified by quantitative autoradiography using tritiated N-methylscopolamine as the ligand. A caudal dorso-medial region is specifically labeled by estrogen receptors identified using immunocytochemistry and by α2-adrenergic receptors which were quantified by autoradiography using tritiated para-amino-clonidine. This latter sub-region is possibly equivalent to the dorso-medial intercollicular nucleus of oscines. Additional track-tracing studies should be performed to confirm this homology. The coexistence of estrogen and α2-adrenergic receptors within the same structure suggests important functional connections between steroid action and catecholaminergic systems in the brain.
    Materialart: Digitale Medien
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  • 89
    Digitale Medien
    Digitale Medien
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 12 (1989), S. 356-363 
    ISSN: 0741-0581
    Schlagwort(e): Adrenal medulla ; Immunocytochemistry ; Light/electron microscopy ; Resinless section ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Allgemeine Naturwissenschaft
    Notizen: In embedment-free electron microscopy with polyethylene glycol embedding and subsequent deembedding, the conventional cytoplasm of the chromaffin cells was revealed to consist of a three-dimensional lattice of microtrabeculae and gives the impression that the chromaffin granules are held in place by the lattice. After the restraint stress, a substantial number of chromaffin cells were almost free of granules, and the microtrabecular lattice was much more compact than that in cytoplasmic regions occupied with remaining granules or increased mitochondria. In immunocytochemistry, actin immunofluorescence was confined to the subplasmalemmal regions, while tubulin and tropomyosin immunofluorescence appeared throughout the entire cytoplasm of normal chromaffin cells. After the stress, the immunofluorescence for actin and tubulin increased in intensity, while that for tropomyosin decreased. Immunogold labelings for actin and tubulin were found mainly on the thinner subplasmalemmal microtrabeculae and the thicker perikaryal ones, respectively, while some were deposited in the form of small aggregates on portions of microtrabeculae. No specific association between the gold labelings for actin or tubulin and the chromaffin granules was found, even in the subplasmalemmal regions. A hypothetical interpretation was proposed in which a more compact lattice of the microtrabeculae in spatial association with a looser lattice represents a gelated state of the cytoplasm. The significance of the gel - sol transition of the cytoplasmic matrix in relation to the secretory mechanism was discussed.
    Zusätzliches Material: 13 Ill.
    Materialart: Digitale Medien
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  • 90
    ISSN: 1432-2307
    Schlagwort(e): AVP/SV40 transgenic mice ; Immunocytochemistry ; Insulin ; Pancreatic polypeptide ; Large T-antigen ; Heterochromatin ; Dysplasia ; Neoplasia
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Insertion into the mouse genome of the hybrid oncogene made up of bovine vasopressin gene derived 5′ upstream sequences and the coding sequences of SV40 large T-antigen promoted tumours in anterior pituitary and endocrine pancreas of mice bearing this transgene. In order to investigate the morphology of the steps in the neoplastic process, we used light and electron microscopy to study these organs in 42 animals belonging to the 3rd, 4th and 5th generations, subdivided into 4 age groups from 20 days to 100 days of life. Antibodies to large T-antigen were used to identify sites of expression of the hybrid oncogene, thus monitoring the steps in neoplastic transformation. Large T-antigen immunoreactivity was identified in dysplastic lesions of younger animals and in both dysplastic lesions and tumours of older mice. Insulin (100% of cases) and pancreatic polypeptide (25% of cases) immunoreactivities were revealed in pancreatic lesions but no hormonal immunoreactivity was detected in the pituitary lesions. The ultrastructural study confirmed that the majority cell population of the pancreatic neoplasms was B-type and that the anterior pituitary tumours were poorly granulated. The subcellular localization of large T-antigen immunoreactivity was investigated by the immunogold method and was confined to the heterochromatin of tumour cell nuclei. These findings provide evidence for the dysplasia-neoplasia sequence in the genesis of endocrine tumours of pituitary and pancreas of transgenic mice. The vasopressin-SV40 large T-antigen transgenic mice may therefore be an useful model for the study of endocrine cell oncogenesis,
    Materialart: Digitale Medien
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  • 91
    ISSN: 1432-2307
    Schlagwort(e): Bartter's syndrome ; Hyperprostaglandin E-syndrome ; Juxtaglomerular apparatus ; Renin-angiotensin system ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary A comparative immunocytochemical and electron microscopic study was performed on renal biopsies from two children with classical Bartter's syndrome (BS) and three children with a recently described variant, the so-called hyperprostaglandin E-syndrome (HES). Compared to age-matched controls, kidney specimens from patients with BS and HES disclosed a marked hypertrophy and hyperplasia of the juxtaglomerular apparatus (JGA). In addition, in HES focal tubular and interstitial calcifications accompanied by interstitial fibrosis and tubular atrophy were noted. On immunocytochemistry, chronic stimulation of the JGA in BS and HES was characterized by an increase in the number of renin-positive cells, particularly in the media of afferent arterioles, but also in efferent arterioles and in the glomerular stalk. The length of the renin-positive portion of the preglomerular arterioles was significantly increased when compared to controls (100±32 vs. 49±17 µm;p〈0.001). In addition, the immunoreactivity of individual renin-positive cells was markedly enhanced. On electron microscopy, “hypertrophy” of the RER and of Golgi complexes with paracrystalline deposits in dilated RER cisterns and protogranules indicated an increased renin synthesis. Renin could be identified in mature secretory granules as well as protogranules by immune electron microscopy. Angiotensinogen was present in hypertrophied epithelial cells of Bowman's capsule. Converting-enzyme reactivity was observed in controls as well as in BS and HES in the brush border of the proximal tubule. In contrast to previous reports, Angiotensin II was completely negative in control as well as in diseased kidneys. We conclude from our results that both BS and HES are characterized by a marked activation of the JGA and severe stimulation of the renin-angiotensin system. Since activation of this system, however, leads - independently of the primary stimulus - to qualitatively very similar morphological reactions, these results do not implicate a common pathogenetic mechanism to both conditions.
    Materialart: Digitale Medien
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  • 92
    ISSN: 1432-2307
    Schlagwort(e): Histopathology ; Immunocytochemistry ; Phaeochromocytoma ; Adrenal gland ; Neuroendocrine markers
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary In this study, antibodies to a range of markers of neuroendocrine differentiation were evaluated for their use in the histopathological assessment and characterisation of phaeochromocytomas. Routinely processed wax blocks from eleven adrenal phaeochromocytomas (10 benign and 1 malignant) and one benign phaeochromocytoma of the urinary bladder were investigated. In addition to these tumours, normal human, cat and piglet adrenal glands were examined. In the phaeochromocytomas, immunostaining was obtained with 21 of the 25 antisera used. Of the general neuroendocrine markers, neuron-specific enolase was found in all tumours, and chromogranin and protein gene-product 9.5 in most of the cases. A range of regulatory peptide immunoreactivities could be demonstrated, such as enkephalin, neuropeptide tyrosine (NPY), 7B2, galanin and vasoactive intestinal polypeptide (VIP). In addition, two peptides were found which have not been reported previously in these tumours, peptide histidine methionine (PHM) and the cryptic fragment of the precursor encoding VIP. Co-localisation studies revealed that peptides derived from the same precursor or peptide family were found in the same tumour cells (e.g. VIP and PHM, NPY and its C-flanking peptide CPON). In the normal adrenal medulla, all the peptides previously reported to be present could be demonstrated immunocytochemically. Galanin was present in a subpopulation of cells also immunoreactive for enkephalin. Neuropeptide tyrosine and CPON were demonstrated in another subpopulation. Occasionally, cells were found to contain all four antigen immunoreactivities. Using antisera to enzymes involved in catecholamine synthesis, galanin was found to be present in noradrenalin-containing cells. The study demonstrates the presence of various antigens in chromaffin tissue of the adrenal gland. A range of substances can also be identified immunocytochemically in phaeochromocytoma tissue, using routinely-processed material.
    Materialart: Digitale Medien
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  • 93
    ISSN: 1432-2307
    Schlagwort(e): Cytokeratins ; Vimentin ; Immunocytochemistry ; Ovarian epithelial tumours ; Intermediate filament coexpression
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary An immunocytochemical investigation has been performed on 83 common epithelial tumours of the ovary, to ascertain their capability of expressing vimentin in addition to cytokeratins. Our results demonstrate that vimentin coexpression is related to the tumour histotype and -to a lesser extent- to the degree of differentiation of malignant variants. Indeed, most serous tumours (80%), some endometrioid adenocarcinomas, and all the clear cell carcinomas investigated exhibited a variable number of neoplastic cells co-synthesizing the two distinct intermediate filament (IF) proteins, whereas only one of 29 mucinous tumours and none of the Brenner tumours displayed vimentin-immunoreactive cells. Moreover, in serous and endometrioid carcinomas, the expression of vimentin was related to the degree of tumour differentiation, being consistently identifiable in the better differentiated cases. The immunocytochemical findings of a parallel investigation on IF expression in the ovarian coelomic epithelium and in the müllerian-derived epithelia of the female genital tract allowed us to ascertain that ovarian epithelial tumours (with the possible exception of poorly differentiated carcinomas) maintain the pattern of IF expression typical of the normal epithelia. This investigation emphasizes the usefulness of IF typing as a tool for the more precise characterization of the origin and differentiation of human neoplasms.
    Materialart: Digitale Medien
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  • 94
    ISSN: 1432-2307
    Schlagwort(e): Herpes simplex virus (HSV) ; Encephalitis ; Immunocytochemistry ; DNA hybridization ; In situ hybridization
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Formol-fixed and paraffin-embedded brain tissue of 33 cases of human necrotizing encephalitis was investigated for Herpes simplex virus (HSV) by immunocytochemistry with a polyclonal antiserum, and by in situ hybridization (ISH) with a biotinylated cDNA probe. HSV antigens (VA) were found in various types of cells in the cytoplasm, cellular processes and nuclei. Labelling by ISH was mostly restricted to nuclei and intranuclear inclusions but otherwise matched the distribution of VA. Eighteen of 25 acute cases had HSV antigen detectable by immunocytochemistry, and 18 of the acute cases contained HSV DNA detectable by our ISH technique. However, results differed somewhat between the techniques: three brains negative for VA showed hybridization, and other 3 VA-positive cases remained negative by ISH. Thus 21 brains with acute necrotizing encephalitis were labelled with one or both techniques. In 8 cases with a subacute course (duration of disease was longer than 1 month), HSV antigens were never detectable although 4 brains showed hybridization. All brains labelled by one or both techniques contained nuclear inclusions bodies. Only one case, of subacute course, with inclusion bodies remained unlabelled. Brain tissue of 11 controls, including cytomegalic inclusion body disease, was never labelled. These results demonstrate that immunocytochemistry and ISH are techniques of comparable sensitivity (72%) for detection of HSV in paraffin sections of acute necrotizing encephalitis brains; their combined use enhanced sensitivity, in our hands, to 84%. In cases with a disease course longer than one month, ISH seems to be the method of choice to demonstrate HSV in situ.
    Materialart: Digitale Medien
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  • 95
    Digitale Medien
    Digitale Medien
    Springer
    Acta neuropathologica 75 (1988), S. 272-276 
    ISSN: 1432-0533
    Schlagwort(e): Alzheimer's disease ; Astrocytes ; Glial fibrillary acidic protein ; Immunocytochemistry ; RNA
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary A new double-staining procedure, in which the techniques of immunocytochemistry of glial fibrillary acidic protein (GFAP) and quantitative microdensitometry of azure B-RNA were combined, was used to study nucleic acid alterations in fibrous astrocytes in Alzheimer's disease (AD). RNA contents of GFAP-positive cells of the hippocampal endplate (Rose's H3-H5 fields) and the dentate gyrus molecular layer were determined in ten autopsy-proven AD patients (ages 51–88) and ten age-matched, non-demented control. In addition, RNA contents of pyramidal neurons of the endplate were examined. While there were no differences in RNA contents of astrocytes of either region between AD patients and controls, neuronal RNA was markedly depleted. These data suggest that astrocytes maintain protein synthetic capabilities in AD and that RNA loss is limited to the neuronal compartment.
    Materialart: Digitale Medien
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  • 96
    Digitale Medien
    Digitale Medien
    Springer
    Acta neuropathologica 77 (1988), S. 91-94 
    ISSN: 1432-0533
    Schlagwort(e): Chromatolytic neurons ; Werdnig-Hoffmann disease ; Immunocytochemistry ; Phosphorylated neurofilament ; Sudden infant death syndrome (SIDS)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Two cases of Werdnig-Hoffman disease are described. Chromatolytic neurons were found within the ventral horns of the spinal cord as well as in dorsal root ganglia, Clarke's column, cranial nerve nuclei and thalamus. Immunohistochemical staining demonstrated ohosphorylated neurofilament antigen within the cytoplasm of the chromatolytic neurons. This finding suggests that a defect in slow axonal transport or in the regulation of neurofilament phosphorylation may play a role in the pathogenesis of this disorder.
    Materialart: Digitale Medien
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  • 97
    Digitale Medien
    Digitale Medien
    Springer
    Acta neuropathologica 77 (1988), S. 168-174 
    ISSN: 1432-0533
    Schlagwort(e): Hirano body ; Immunocytochemistry ; Actin ; Microtubule-associated proteins ; Tropomyosin
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary Hirano bodies are filamentous, paracrystalline inclusions that are found in dendrites and cell bodies of neurons in Alzheimer's and other neurodegenerative diseases. Actin appears to be a major component of these structures. We present evidence that tropomyosin and high molecular weight microtubule-associated proteins (MAPs) are also components of Hirano bodies. Although an association betwen actin and MAPs has been noted in vitro, interactions in vivo have not heretofore been demonstrated. Since microtubules are not present in Hirano bodies, and anti-tubulin and anti-neurofilament antibodies do not bind to Hirano bodies, the association between MAPs and these inclusions is likely a result of interactions between MAPs and actin, and not MAPs and microtubules or neurofilaments.
    Materialart: Digitale Medien
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  • 98
    Digitale Medien
    Digitale Medien
    Springer
    Anatomy and embryology 178 (1988), S. 137-142 
    ISSN: 1432-0568
    Schlagwort(e): Endothelium ; Blood vessels ; 5-Hydroxytryptamine ; Substance P ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary An electron microscopic-immunocytochemical study has been made on the localisation of serotonin and substance P in endothelial cells of femoral and mesenteric arteries of the rat. In the femoral and mesenteric arteries, less than 10% of the endothelial cells showed positive immunolabelling for these substances. The distribution of serotonin and substance P in endothelial cells was similar in both arteries. The immunoprecipitate was confined to the cytoplasm, including subcellular organelles. The results are discussed in terms of endothelial mechanisms of local control of blood flow.
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  • 99
    Digitale Medien
    Digitale Medien
    Springer
    Anatomy and embryology 179 (1988), S. 165-172 
    ISSN: 1432-0568
    Schlagwort(e): Somatostatin-14 ; Human olfactory bulb ; Anterior olfactory nucleus ; Immunocytochemistry
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary This study describes the morphological features and the distribution pattern of neurons in the human olfactory bulb which are immunoreactive for an antiserum against the neuropeptide somatostatin-14. Immunoreactive nerve cell bodies were mainly found in the white matter surrounding the cell clusters of the anterior olfactory nucleus. Some immunoreactive neurons were also found scattered throughout the anterior olfactory nucleus and the deeper parts of the inner granule cell layer. Only a few immunoreactive neurons were localized in the glomerular layer and the outer granule cell layer. Immunoreactive fibres were found in all layers of the olfactory bulb. In addition, an impressive number of coiled and kinked immunoreactive fibres were localized within the anterior olfactory nucleus forming a dense plexus. Accumulations of twisted and coiled branches of immunoreactive fibres were rarely found either surrounding or within the olfactory glomerula. The characteristics of somatostatin-14 immunoreactive neurons as seen in the combined pigment-Nissl preparation were studied after decolourizing the chromogen and restaining the preparations with aldehydefuchsin in order to demonstrate the lipofuscin pigment and gallocyanin chrome alum for Nissl material. About 90% of the immunoreactive neurons studied in this manner turned out to be devoid of lipofuscin granules. The remaining 10% displayed different patterns of pigmentation. These findings suggest the presence of different types of somatostatin-14-like immunoreactive neurons in the olfactory bulb of the human adult.
    Materialart: Digitale Medien
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  • 100
    Digitale Medien
    Digitale Medien
    Springer
    Acta neuropathologica 75 (1988), S. 345-353 
    ISSN: 1432-0533
    Schlagwort(e): Lewy body ; Immunocytochemistry ; Ubiquitin ; Paired helical filaments
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Summary The nature of Lewy bodies (LBs) in the brain stem and cerebral cortex in five cases of diffuse Lewy body disease and one case of Parkinson's disease with dementia were investigated immunocytochemically with various antibodies to cytoskeletal proteins, paired helical filaments (PHF) and ubiquitin. Antibodies to 200-kDa component of neurofilament, tau and PHF showed no significant reactions with most of LBs. Antibodies to high-molecular weight microtubule-associated proteins (HMWMAPs) moderately stained the periphery of a few of LBs. A monoclonal antibody to PHF (DF2) which recognizes ubiquitin, and polyclonal antibodies to ubiquitin immunostained virtually all of the typical and cortical LBs as intensely as Alzheimer's neurofibrillary tangles and senile plaque neurites: the periphery of LBs was darkly stained, whereas the central core of typical LBs and central zone of cortical LBs were less intensely stained or remained unstained. Immunoelectron microscopy of the LBs with DF2 revealed that immune reaction products were located on the filaments exclusively in the periphery of LBs, but not on those in the center. These findings suggest that both types of LBs are immunocytochemically indistinguishable despite some structural differences, and that peripherally located filaments in LBs are tagged with ubiquitin, an element required for the ATP-dependent proteolysis system in the cell. Antibodies to ubiquitin are the most useful marker of LBs ever known.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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