Frans Vandesande

Identification of the vasopressin producing and of the oxytocin producing neurons in the hypothalamic magnocellular neurosecretory system of the rat

SummaryImmuno-enzyme histochemical investigations showed that, in the magnocellular hypothalamo-hypophysial neurosecretory system of the rat, vasopressin and oxytocin are synthetized in separate neurons. Both the vasopressin neurons and the oxytocin neurons are present in both the supraoptic and the paraventricular nuclei in about the same number. Preferential location of the two kinds of rat neurosecretory neurons is not as obvious as in the bovine hypothalamus. Their perikarya do not show distinct morphological differences. The two kinds of neurosecretory perikarya are the origin of separate vasopressin-containing and oxytocin-containing axons respectively. In the neural lobe, the distribution of the two different types of axons is described.

The origin of the vasopressinergic and oxytocinergic fibres of the external region of the median eminence of the rat hypophysis

SummaryThe origin of the vasopressinergic and oxytocinergic nerve fibres of the external region of the rat median eminence was investigated by means of hypothalamic lesions, adrenalectomy and immunocytochemistry. The results obtained in bilaterally adrenalectomized animals with complete, or incomplete, destruction of the suprachiasmatic nuclei showed that, at least, the great majority of the vasopressinergic and oxytocinergic nerve fibres of the external region of the rat median eminence do not originate from the suprachiasmatic nuclei. From the observations obtained in bilaterally adrenalectomized animals with total or subtotal destruction of both paraventricular hypothalamic nuclei, it appears that the paraventricular nuclei must be the origin of (nearly) all the vasopressinergic and oxytocinergic nerve fibres of the external region of the rat median eminence. The results strongly suggest that both types of fibres originate from all parts of the paraventricular nuclei.

Identification of the vasopressin-neurophysin producing neurons of the rat suprachiasmatic nuclei.

SummaryImmuno-enzyme histochemical investigations on the hypothalamus of the normal rat showed (1) that the Suprachiasmatic nuclei produce vasopressin; (2) that it is highly probable that these nuclei do not produce oxytocin.From the present and previous investigations it may be concluded that the Suprachiasmatic neurons produce a vasopressin-neurophysin complex.

Immunocytochemical localization of somatostatin-containing neurons in the rat hypothalamus.

SummaryThe rat hypothalamus was studied at the light microscopic level with the use of single and double immunocytochemical staining methods. It was shown that the rat supraoptic and paraventricular hypothalamic nuclei, and their accessory neurosecretory nuclei, do not contain magnocellular somatostatin neurons. The distribution of the hypothalamic parvocellular somatostatin cells is described. The parvocellular component of the rat hypothalamic paraventricular nucleus is, at least partly, composed of somatostatin cells: they form a fairly well circumscribed periventricular cell mass. The rat suprachiasmatic nuclei contain separate somatostatin neurons and vasopressin neurons. Scattered somatostatin cells are present in the entire arcuate nucleus. In addition to the periventricular somatostatin cells located in the preopticanterior hypothalamic area and in the arcuate nucleus, the rat hypothalamus also contains numerous scattered somatostatin cells located distant from the third ventricle.

Heterogeneity of GABAergic cells in cat visual cortex

Antibodies against neuropeptides and against a vitamin D-dependent calcium-binding protein (CaBP) label small cells with nonpyramidal-like morphology in the cat visual cortex (areas 17, 18, and 19). Since GABAergic cells are interneurons, a double-staining procedure was used to test for the coexistence of cholecystokinin (CCK), somatostatin (SRIF), neuropeptide Y (NPY), corticotropin-releasing factor (CRF), vasoactive intestinal polypeptide (VIP), and CaBP with glutamic acid decarboxylase (GAD). Our results show that CRF and VIP do not coexist with GAD, while the 3 other peptides and CaBP do. Hence GAD-positive cells can be subdivided into 4 broad groups: (1) cells that are only GAD-positive, (2) cells that are GAD- and CaBP-positive, (3) GAD- positive neurons also containing CCK, and (4) GAD-positive cells that also contain SRIF. A small subset of class 2 also contains SRIF and most cells of class 4 also contain NPY. The 4 classes of GAD-positive cells differ in laminar position: class 1 predominates in layers IV and V, classes 2 and 3 in the upper laminae (II and III), and class 4 in the deepest layer (VI).

Identification of 26RFa, a hypothalamic neuropeptide of the RFamide peptide family with orexigenic activity

A neuropeptide was isolated from a frog brain extract by HPLC purification and characterized by mass spectrometry. This 26-aa neuropeptide, which belongs to the RFamide peptide family, was designated 26RFa, and its primary structure was established as VGTALGSLAEELNGYNRKKGGFSFRF-NH2. Research in databases revealed the presence of sequences homologous to frog 26RFa in the human genome and in rat ESTs. On the basis of this sequence information, the cDNAs encoding the human and rat 26RFa precursors were cloned. The two preproteins show a similar organization, with the 26RFa sequence located in the C-terminal region of the precursor. Human preprotein (prepro)-26RFa encodes an additional putative RFamide peptide that is not found in the rat precursor. The primary structures of human, rat, and frog 26RFa exhibit ≈80% identity, and the C-terminal octapeptide has been fully conserved from amphibians to mammals. In situ hybridization histochemistry revealed that, in the rat brain, the 26RFa gene is exclusively expressed in the ventromedial hypothalamic nucleus and in the lateral hypothalamic area. 26RFa induced a dose-dependent stimulation in cAMP production by rat pituitary cells in vitro and markedly increased food intake in mice. The conservation of the primary structure of 26RFa during vertebrate evolution, the discrete localization of the mRNA encoding its precursor in hypothalamic nuclei involved in the control of feeding behavior, and the observation that 26RFa possesses orexigenic properties indicate that this neuropeptide may play important biological functions.

Immunocytochemical localization of the vasopressinergic and the oxytocinergic neurons in the human hypothalamus

SummaryThe human hypothalamic-neurohypophysial hormone-producing nuclei were investigated with the unlabeled antibody peroxidase-antiperoxidase complex (PAP) technique at the light microscopic level. The size, shape and location of the supraoptic, paraventricular, accesssory supraoptic and suprachiasmatic nuclei were determined. It was demonstrated in the human hypothalamus, as well as in the hypothalamus of other mammals, that vasopressin and oxytocin are synthesized in separate neurons. In each of the nuclei of the magnocellular neurosecretory system, the distribution, ratios and structural features of the vasopressinergic and oxytocinergic neurons were determined. It was shown that the human suprachiasmatic nuclei contain numerous neurophysin-vasopressin-producing neurons.

Identification of the vasopressin-neurophysin II and the oxytocin-neurophysin I producing neurons in the bovine hypothalamus

SummaryImmuno-enzyme histochemical investigations on the bovine hypothalamus showed that vasopressin and oxytocin are synthetized in separate neurons. Moreover, it was found that the vasopressin-producing neurons are identical with the neurophysin II-producing neurons, while the oxytocin-producing neurons correspond with the neurophysin I-producing neurons. From this result it is concluded that, in the species studied, neurophysin I is the carrier protein of oxytocin and that neurophysin II is the carrier protein of vasopressin.The bovine vasopressin-neurophysin II producing neurons and the oxytocin-neurophysin I producing neurons are both present as well in the supraoptic nuclei as in the paraventricular nuclei. In the supraoptic nuclei, the vasopressin-neurophysin II neurons predominate in number; in the paraventricular nuclei, the oxytocin-neurophysin I neurons predominate. The two kinds of bovine neurosecretory neurons are partly localized in separate areas. Moreover, the vasopressin-neurophysin II neurons and the oxytocin-neurophysin I neurons show distinct morphological differences.

CALCIUM BINDING PROTEINS AND NEUROPEPTIDES AS MOLECULAR MARKERS OF GABAERGIC INTERNEURONS IN THE CAT VISUAL CORTEX

SummaryIn the cat visual cortex, almost all parvalbumin-positive cells are GABAergic, and about 80% of the calbindin D-28K-positive neurons are also GABA-immunoreactive. About 37% of the GABAergic neurons contain parvalbumin and a smaller fraction (about 18%) contains calbindin. Furthermore, parvalbumin and calbindin are localized in two separate neuronal populations in the cat visual cortex, suggesting that two GABAergic populations can be distinguished, one containing parvalbumin and one containing calbindin. Double staining for parvalbumin and neuropeptides (CCK, SRIF and NPY), revealed no double-labeled cells, with the exception of a few SRIF- and parvalbumin-positive neurons. These results show that cortical GABAergic cells can be differentiated on basis of their calcium binding protein and neuropeptide immunoreactivity.

Coexistence of cholecystokinin and oxytocin-neurophysin in some magnocellular hypothalamo-hypophyseal neurons

SummaryThe existence of cholecystokinin in the posterior hypophysis and its hypothalamic origin have been unequivocally demonstrated. Immunocytochemical evidence is presented for the coexistence of gastrin-cholecystokinin and oxytocin-neurophysin I immunoreactivities in some magnocellular neurons of the supraoptic and paraventricular nuclei both in rat and bovine hypothalamus.