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  • 1
    ISSN: 1432-1106
    Keywords: Intermediate lobe ; Median eminence ; GABA ; Dopamine ; Colocalization ; Mammal
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Immunoreactivities for tyrosine hydroxylase (TH), gamma-aminobutyric acid (GABA) and, in some cases, glutamic acid decarboxylase (GAD) were detected by light and electron microscopy in axons projecting into the median eminence and pituitary gland of various mammals (rats, mice, guinea pigs, cats, rabbits and hares). Light microscope immunoperoxidase reactions were performed on adjacent semithin sections of plasticembedded samples. In the median eminence external zone, the distributions of the TH- and GAD- or GABA-immunoreactive endings were very similar in the anterior and lateral areas, while medially the GABA-labelled endings predominated. Comparable distribution patterns were found in the various species examined. In the pituitary gland, the distributions of GABA- and TH- immunoreactivities were superimposable in the intermediate lobes of all species examined, except in the rabbit and hare in which both types of innervation were lacking. For electron microscopy, the immunogold procedure was applied to sections of lowicryl-embedded samples; simultaneous detection of GABA- and TH-immunoreactivities was enabled by recto-verso double labelling with gold particles of distinct diameters. In the median eminence, GABA-immunoreactivity occurred systematically in the TH-positive endings, while distinct GABA-positive/TH-negative axons were also detected. In the intermediate lobe, the colocalization of TH- and GABA-immunoreactivities was a constant feature of the axons innervating the melanotrophic cells in all the species examined, except in the Leporidae. The functional significance of this colocalization remains to be determined.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0878
    Keywords: Fetal intermediate lobe ; Tissue culture ; Immunocytochemistry ; In situ hybridization ; Dopamine ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Organotypic cultures, in defined medium, of pituitary primordia obtained from 15-day-old rat fetuses were performed in order to study the in vitro differentiation of melanotrophic cells. The morphological and ultrastructural features of the transplants resembled those of the gland developing in vivo. In situ hybridization on semi-thin sections, using a 35S-labelled oligonucleotide probe, revealed pro-opiomelanocortin-mRNA-containing cells on the first day of culture in the anterior lobe and after 2–3 days in the intermediate lobe. Immunoperoxidase labelling of adjacent sections showed that the same cells reacted with antibodies against α-melanocyte-stimulating hormone (αMSH), γ3 and adrenocorticotropic hormone in both lobes. The pro-opiomelanocortin-mRNA-containing cells formed progressively conspicuous areas in the intermediate lobe, which was almost uniformly labelled after 6 days. In the anterior lobe, these cells remained scattered in small cell groups, and colloidal gold immunolabelling showed the progressive disappearance of αMSH labelling from the secretory vesicles in cells exhibiting morphological features of adult corticotrophic cells. Both the αMSH content of the explants and αMSH release into the culture medium increased with time. Treatment with the dopamine agonist bromocriptine induced a strong dose-dependent decrease in αMSH secretion, which was significant after 3 days in culture, indicating that dopamine D2 receptors are able to regulate hormonal release of melanotrophic cells at early stages. This system constitutes a suitable model for further studies of factors controlling cell differentiation and cellular interactions involved in histogenesis.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0878
    Keywords: Pars intermedia ; Oxytocin ; Dopamine ; Serotonin ; Immunocytochemistry ; Rabbit, hare
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The pars intermedia of the pituitary in the rabbit and hare is abundantly innervated by axons reacting selectively with antibodies against oxytocin. These axons contain dense secretory vesicles about 140 nm in diameter, i.e., smaller than those in the neurosecretory axons of the neural lobe. No fiber elements staining for other peptides (vasopressin, somatostatin, substance P) were observed in the pars intermedia, except rare leu-enkephalin axons restricted to the rostral zone of the gland. Dopaminergic innervation appears to be completely absent from the intermediate lobe. This was shown by the lack of reaction with an antibody against tyrosine-hydroxylase, which did reveal a well-developed tubero-infundibular system of nerve fibers. Axons reacting with an antibody against serotonin were irregularly distributed in the pars intermedia. In the absence of dopaminergic axons, the extensive oxytocin-like innervation may play a major role in regulating the melanotrophic cell activity in the Leporidae.
    Type of Medium: Electronic Resource
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