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  • 1
    ISSN: 1432-0568
    Keywords: HNK-1 ; Heart conduction system ; Bisdiamine ; Rat embryo ; Computer graphics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The spatiotemporal distribution of the immunoreactivity of monoclonal antibody HNK-1 was investigated immunohistochemically in normal and bis-diamine-induced malformed rat embryonic hearts using three-dimensional reconstruction with computer graphics. First recognized in the primitive heart 11.5 days after conception, HNK-1 immunoreactivity was distributed in the atrio-ventricular and bulbo-ventricular junctional areas with incomplete ring-like appearance in the early embryonic stages. In the late embryonic stages the immunoreactive sites were rearranged and localized in the sites topographically corresponding to almost the entire pathway of the conduction system, including the three major internodal tracts connecting the right sinoatrial node and atrioventricular node. Immunoreactivity gradually decreased after the completion of the conduction system, and only a faint reactivity in the atrio-ventricular node region remained in the new-born heart. These results indicate that HNK-1 is expressed temporarily in the pathways corresponding to the conduction system during the development of the heart. In bis-(dichloro-acethyl)-octamethylen-diamine (bis-diamine)-induced malformed hearts, localization of HNK-1 immunoreactivity was not remarkably altered in the early embryonic heart. In the late embryo, immunoreactive sites in the sino-atrial node region and atrio-ventricular node region deviated dorsocaudally with the poorly developed internodal tracts, and abnormal distribution was observed in the bilateral atria. We consider that these abnormalities may occur in conjunction with abnormal morphological development such as insufficient absorption of the sinus venosus.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0568
    Keywords: HNK-1 ; Immunoelectron microscopy ; Conduction system ; Embryo
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To confirm the role of HNK-1 in conduction tissue, the ultrastructural localization of monoclonal antibody HNK-1 was analyzed in developing rat hearts at embryonal day 14.5 by immunoelectron microscopic labeling procedures with post-embedding immunogold staining. Tissue sections in different planes containing the sino-atrial (SA) node, atrio-ventricular (AV) node and His bundle were used to demonstrate HNK-1. Immunogold labeling was detected on the cell surfaces and in the extracellular matrices of cells that had features common to conduction tissue cells. Non-specialized contractile myocytes were not labeled by this antibody. Furthermore, immunogold labeling was more prominent in wide intercellular spaces than in narrow intercellular spaces, and rarely observed in cell-cell contact regions. The cell surfaces and extracellular matrices of mesenchymal cells in the endocardial cushion, which contacts the His bundle, were also positive, suggesting the involvement of tract formation to the AV node. These findings may indicate that HNK-1 plays an important role in cell-cell adhesion processes both temporally and spatially in the developing conduction tissue. It was concluded, therefore, that HNK-1 is a suitable marker of the embryonic heart conduction system and might be useful in analyzing anomalous conduction systems, as in congenital heart disease.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0568
    Keywords: Acetylcholinesterase ; HNK-1 ; Heart ; Morphogenesis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Acetylcholinesterase (AChE) activity was topographically investigated in the presumptive cardiac conduction tissue regions visualized by HNK-1 immunoreactivity in rat embryos, and AChE-positive cells were examined with the electron microscope. On embryonic day (ED) 14.5, when HNK-1 was most intensely visualized, AChE activity could not be detected enzyme-histochemically in the conduction tissue regions, except in the ventricular trabeculae and part of the AV node. On ED 16.5, however, the AChE activity was clearly demonstrated in some parts of the developing conduction tissue. One exception was the AV node region, where an AChE-positive area was in close proximity to an area showing HNK-1 immunoreactivity but did not overlap. Furthermore, AChE activity was demonstrated predominantly in the ventricular trabeculae, including cardiac myocytes, but was rather weak in the atrium. With the electron microscope, AChE reaction products were observed predominantly intracellulary in both developing conduction tissue cells and developing ordinary myocytes, and no reactivity was found in neuronal components. From ED 18.5 until birth, both AChE activity and HNK-1 immunoreactivity faded away in the conduction tissue. Thus, transient AChE activity in the embryonic heart seems to be different from the developing adult form and may be related to a morphogenetic function in embryonic tissues, as proposed by other authors.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0568
    Keywords: Immunohistochemistry ; Leu-7 ; Conduction system ; Embryo
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The distribution pattern of Leu-7 (HNK-1) in developing human embryonic hearts and rat hearts was studied by immunohistochemistry. Human and rat embryos at Streeter's stages XIII ∼ XX and fetus stage I were used. Leu-7, which is absent in the newborn rat heart, is expressed transiently in the embryo and fetus I stages. The earliest embryonic heart shows two incomplete circular structures with immunoreactivity in the myocardium along the primitive atrioventricular cushion and bulboventricular canal. These two structures become localized topographically in the definitive atrioventricular node and atrioventricular bundle after rearrangement and partial disappearance during embryonic development. At Streeter's stages XVIII ∼ XX, Leu-7 immunoreactivity appears to localize topographically in almost all the pathways of the conduction system, although some discontinuities are observed in the atrioventricular junction and atrial internodal tracts. Thereafter, immunoreactivity decreases gradually and differentially by site and stage. The precise nature of Leu-7 immunoreactive cells, that is, whether or not they are neurogenic or myogenic, is not revealed by this study. The present observations are discussed in connection with the hypothesis that specialized ring tissue is the primordium of the conduction system.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1436-2813
    Keywords: Key Words: malignant mesothelioma ; tunica vaginalis testis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Cellular and molecular neurobiology 17 (1997), S. 141-150 
    ISSN: 1573-6830
    Keywords: vascular endothelial growth factor ; basic fibroblast growth factor ; ethylnitrosourea-induced rat glioma ; digoxigenin ; in situ hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 1. Which angiogenic growth factors actually mediate tumor growth in ethylnitrosourea (ENU)-induced gliomas in rats was examined. 2. In situ hybridization histochemistry with digoxigenin-labeled oligonucleotide probes was used to investigate the cellular expression and distribution of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) mRNAs in ENU-induced gliomas. 3. Both VEGF and bFGF mRNAs were not detected in normal gial cells but in ENU-induced glioma cells. 4. Our results suggest that the growth of ENU-induced glioma may be regulated by multiple angiogenic growth factors and that these gliomas may proliferate by synthesizing such growth factors.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-7373
    Keywords: ethylnitrosourea ; glioma ; cell line ; GFAP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In order to evaluate the proliferation and differentiation potentials of ethylnitrosourea (ENU)-induced rat glioma cells, the authors attempted to obtain a cell line that maintains glial features in long-term culture. One of five cell lines cultivated from ENU-induced rat gliomas merited particular interest because of the differentiation of its neoplastic glia. This cell line, designated as HITS glioma, had a polygonal cell body and formed a monolayer with pile-up fociin vitro, in contrast to the other cell lines, which displayed a mesenchymal change through passages. GFAP-positive cells, found in the primary culture, disappeared in the late passages of HITS glioma, as they did in the other cell lines. Galactocerebroside (GC), GD3 ganglioside, and Leu7 were not expressed in the cell lines during culture. Subcutaneous inoculation of HITS glioma into neonatal rats induced tumors with histopathological components mimicking the histopathological appearance of ENU-induced gliomas. The components also had a fraction of GFAP-positive cells. Such findings indicate that HITS glioma cells may be composed of immature glial cells which are able to differentiate into astrocytic cells under certain conditions. Several growth factors which play a role in gliogenesis were used to evaluate the mechanism(s) of proliferation and/or differentiation of HITS glioma. These growth factors did not induce the expression of GFAP and other antigenic expression in HITS glioma, even though some promoted the proliferation of HITS glioma. Although the mechanism involving the astrocytic differentiation of HITS glioma is unknown, HITS glioma may serve as an effective tool in research to evaluate the mechanisms of proliferation and differentiation of neoplastic glia.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-0878
    Keywords: Key words: Aging ; Dietary restriction ; Hepatocytes ; Cell proliferation ; Proliferating cell nuclear antigen ; Cell death ; Terminal dUTP nick end labeling (TUNEL) ; Rat (Fischer 344)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The proliferation and death of hepatocytes in rats fed ad libitum and rats on dietary restriction were evaluated in 3 to 24-month-old rats by employing immunocytochemistry for proliferating cell nuclear antigen (PCNA) and terminal dUTP nick end labeling (TUNEL). These techniques were also used to examine hepatic tissue infiltrated with leukemic cells in 24-month-old rats fed ad libitum. PCNA-strongly positive hepatocytes, PCNA-positive hepatocytes, and TUNEL-positive hepatocytes were reported previously to be equivalent to hepatocytes in the S phase, hepatocytes in the cell cycle, and dying hepatocytes, respectively. The proportion of PCNA-strongly positive hepatocytes and PCNA-positive hepatocytes declined with age. Dietary restriction diminished PCNA-strongly positive hepatocytes significantly but not PCNA-positive hepatocytes in young rats, but the proportion of PCNA-strongly positive hepatocytes was significantly higher following dietary restriction than that in rats fed ad libitum in advanced age. Growth stimulation by leukemic cell infiltration resulted in a recovery of the age-related decline of PCNA-strongly positive hepatocytes. Aging was associated with a progressive increase in the proportion of TUNEL-positive hepatocytes, with a smaller effect following dietary restriction than in rats fed ad libitum after 6 months of age. Our results indicate that age and dietary restriction induce proliferative inhibition. The inhibition depends on PCNA expression; this suggests that suppression of cell proliferation and cell death are enhanced in hepatocytes of senile rats.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1574-4647
    Keywords: Aging ; dietary restriction ; somatotrope ; cell renewal ; growth hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We investigated the impact of dietary restriction on the basal rate of somatotrope renewal in the pituitary gland. Bromodeoxyuridine (BrdU), a thymidine analog, was administered continuously for 1 week in male F344 rats at 3, 8 and 20 months of age (mo), fed ad libitum (AL) or diet restricted from 1.5 mo (DR). Combined immunostainings for BrdU and GH visualized newly formed somatotropes as well as pituitary cells in tissue sections. The rate of incorporation of BrdU by anterior pituitary cells (BrdU-labeled nuclei/100 nuclei) was not influenced by the dietary regimen or age. The fraction of BrdU-labeled somatotropes relative to all labeled cells precipitously decreased to the same level in both dietary groups between 3 and 8 mo, although the fraction was greater in DR rats at 3 mo. In AL rats, the fraction decreased further between 8 and 20 mo, while it stabilized in DR rats. Our results suggested that dietary restriction maintains the basal rate of somatotrope renewal in later life in male rats. Although one must also estimate the effects of dietary restriction on apoptotic cell death in pituitary cells, the present study provides evidence that dietary restriction modulates somatotropes cell turnover and preserves the cell population for GH secretion during aging.
    Type of Medium: Electronic Resource
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