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  • 1
    ISSN: 1432-0878
    Keywords: Somatotropes, growth hormone cells ; Immunocytochemistry ; Growth hormone (GH) ; Receptors, membrane ; Somatostatin (SRIF) ; Growth hormone-releasing hormone (GRH) ; Rat (Han: WIST)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: 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.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0878
    Keywords: Somatostatin ; Cholecystokinin ; Binding sites ; Receptors ; Neurons ; Telencephalon ; Rat (Han: WIST)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The identification of high-affinity binding sites for neuropeptides on individual target cells is a prerequisite when studying the sites of action and the manner in which peptides act as neuromediators. In situ and in vitro, this can be achieved using newly synthesized, biologically active conjugates of somatostatin or cholecystokinin (sulphated octapeptide) with colloidal gold. Labelled neurons show a peptide-specific, non-overlapping distribution in rat telencephalic structures; i.e, whereas the somatostatin-gold conjugate labels binding sites on neurons and glial cells, cholecystokinin-binding sites are restricted to neurons. Binding of either gold-labelled ligand can be competitively suppressed by excess amounts of the native peptide or its analogues. Neuronal somatostatin-binding sites are visualized on neurons in lamina III and, in particular, in lamina V/VI of the primary somatosensory cortex and in the magnocellular nucleus of the telencephalic cholinergic system. Cholecystokinin-binding sites are localized in the main olfactory bulb, on neurons in the cortical “hindlimb” and “forelimb” region, in the hippocampus, and in the cingulate and visual cortex.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 262 (1990), S. 431-443 
    ISSN: 1432-0878
    Keywords: Astrocytes ; Telencephalon ; Receptors, membrane ; Somatostatin (SRIF) ; Rat (Han: WIST)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using a somatostatin-gold conjugate as ligand, high-affinity binding sites for this neuropeptide were demonstrated at three levels: (i) cultured astrocytes from rat cortex, (ii) hippocampal slice cultures, and (iii) frozen tissue sections of rat telencephalon. The conjugate proved as active as the native peptide in competing for the binding sites. Light-microscopic visualization of bound ligand was achieved by silver intensification of the colloidal gold. This method is faster and yields superior resolution compared with autoradiography. Cultured astrocytes from cortex and hippocampus could be labeled by the ligand. At the light- and electron-microscopic level, astrocytes could be double-labeled by the somatostatin-gold conjugate and immunostaining for glial fibrillary acidic protein (GFAP). In hippocampal slice cultures, the conjugate did not penetrate into the neuropil because of a covering glial layer. However, a portion of this completely GFAP-positive covering glia reacted with the somatostatin ligand. In frozen brain sections, apart from delicate punctate structures, two types of labeled glia cells were seen: single stellate astrocytes and perivascular glia cells.
    Type of Medium: Electronic Resource
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