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  • 1
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Electron microscope immunocytochemistry was used to determine the intracellular localization and distribution among follicular elements of four peptides: calcitonin, somatostatin, calcitonin gene-related peptide (CGRP), and substance P in the thyroid glands of bats captured in the prehibernation phase of their annual life cycle. Previous studies have shown that this period of the hibernation - activity cycle is characterized by the accumulation and storage of secretory granules in parafollicular cells. Sites of binding of primary antisera to each of the four peptides were identified by means of affinity-purified secondary antisera directly coupled to colloidal gold particles. Calcitonin and somatostatin immunoreactivities were found in all parafollicular cells examined and in every secretory granule within these cells. CGRP was also found in all parafollicular cells examined (n = 75) but only in about half of their secretory granules. In contrast to these peptides, substance P immunoreactivity was not found in any parafollicular cells, but was localized exclusively in nerve endings within the basement membrane of the follicle.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 226 (1990), S. 471-480 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The distribution and cellular localization of the glycoprotein laminin were investigated by light and electron microscopic immunocytochemistry in the adult murine pituitary gland. Immunoblots confirmed that laminin was the only protein in the pituitary gland of the adult male mouse to react with antilaminin serum. Laminin immunoreactivity was demonstrated at the light microscopic level simultaneously with that of β-follicle stimulating hormone (β-FSH) and β-luteinizing hormone (β-LH). In addition to its distribution is basal laminae, laminin immunoreactivity was coincidently expressed in gonadotrophs with the immunoreactivities of β-FSH and β-LH. Electron microscopic immunocytochemistry was employed on aldehyde-fixed sections embedded in L.R. White. Sites of binding of primary antisera to laminin were identified with affinity-purified secondary antisera directly coupled to 20 nm particles of colloidal gold. Three antisera recognizing laminin were compared and found to result in an identical pattern of immunoreactivity. Laminin was found extracellulary only in formed basal laminae in all three lobes of the pituitary and was not found in extracellular matrices of connective tissue. Laminin immunoreactivity was also found intracellularly in gonadotrophs but in none of the other endocrine or non-endocrine cells of the anterior lobe. Within gonadotrophs, only secretory granules were labeled. The majority, but not all, secretory granules were labeled in each of the gonadotrophs examined, and the proportion of granules labeled with laminin could not be increased by doubling the concentration of anti-laminin serum. Laminin immunocreactivity segregated with the subset of secretory granules containing β-FSH. In contrast, laminin immunoreactivity was absent in the smaller subset of secretory granules that contain serotonin. No secretory granules were labeled within the endocrine cells of the intermediate lobe, nor within secretory granules of neural elements in the posterior lobe.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 215 (1986), S. 51-58 
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Previous studies have demonstrated the uptake of exogenous and storage of endogenous 5-hydroxytryptamine (5-HT) in parenchymal cells of the anterior pituitary. The present experiments were undertaken to test the hypothesis that the endogenous 5-HT of the anterior lobe of the pituitary is costored with β-luteinizing hormone (β-LH) within the same secretory granules of gonadotrophs. Electron microscope immunocytochemistry was used to detect 5-HT and β-LH immunoreactivities in the anterior pituitary glands of mice and bats. Primary antisera generated in different species of animals to these two antigens were localized with appropriate species-specific secondary antisera coupled to colloidal gold particles of different sizes. This enabled 5-HT and β-LH immunoreactivities to be demonstrated simultaneously on ultrathin sections of fixed anterior lobe tissue mounted on electron microscope (EM) grids. In both bats and mice 5-HT immunoreactivity, identified by immunostaining of β-LH, was found in gonadotrophs, and in no other cell type. Within gonadotrophs about 25% of the secretory granules were labeled by antisera to both 5-HT and β-LH, although 100% of granules reacted with the antiserum to B-LH. No secretory granules were found that were immunostained only by the antiserum to 5-HT. It is concluded that endogenous 5-HT may be a normal constituent of mammalian gonadotrophs and that it is colocalized with β-LH in at least a subset of the secretory granules of these cells. It cannot yet be concluded that gonadotrophs synthesize 5-HT as well as taking it up from the ambient medium. The function of 5-HT in the pituitary remains unknown; however, the costorage of 5-HT and β-LH in the same secretory granules would seem to be a mechanism to ensure the simultaneous release of the two substances by exocytosis.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The hypothesis that the secretory granules of mammalian gonadotrophs are heterogeneous was tested. Previous studies had shown that all of the granules contain β-luteinizing hormone (β-LH) immunoreactivity, and some contain β-follicle stimulating hormone (β-FSH) or 5-HT immunoreactivity. Moreover, differential release of β-LH and β-FSH has also been demonstrated. In the current study the pituitary glands of mice were investigated immunocytochemically at the ultrastructural level with antisera directed against human growth hormone (GH), β-LH, β-FSH, and 5-HT. The immunoreactivities of β-LH, β-FSH, and 5-HT were restricted to gonadotrophs. No 5-HT immunoreactivity was seen in somatotrophs, identified by the immunoreactivity of GH in the secretory granules of these cells. Antisera to β-LH labeled all gonadotroph granules; however, anti-β-FSH and anti-5-HT sera labeled only subsets of the granules. The proportion of granules labeled could not be increased by doubling the concentration of anti-β-FSH serum. The incidence of double labeling of granules by antisera to β-FSH and 5-HT was significantly less than that predicted from the incidence of granule labeling by either reagent alone. It is concluded that β-FSH and 5-HT immunoreactivities do not co-exist in the same secretory granules of gonadotrophs; therefore, these granules are heterogeneous and there must be at least two types of granules. It is possible that the two types of granules may be responsive to different second messengers, thereby explaining the differential release of LH and FSH.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
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