Peptide‐ and transmitter‐containing neurons in the mediobasal hypothalamus and their relation to GABAergic systems: Possible roles in control of prolactin and growth hormone secretion

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Abstract
Indirect immunofluorescence histochemistry was used to study the relation among GABAergic, catecholaminergic, cholinergic, and peptidergic neurons in the rat mediobasal hypothalamus. By employing a direct double‐labelling procedure using sheep antiserum against glutamic acid decarboxylase (GAD), mouse monoclonal and rabbit antibodies to neurotensin (NT) and rabbit antisera to tyrosine hydroxylase (TH),choline acetyltransferase (ChAT), galanin (GAL), growth hormone‐releasing factor (GRF), or somatostatin (SOM), it was demonstrated that GAD‐positive fibers and terminals in the external part of the median eminence co‐contained immunoreactivity for TH, NT, GAL or GRF, but not for SOM. In the internal part of the median eminence‐infundibular stalk, GAD‐positive/NT‐, GAL‐, and GRF‐negative and GAD‐Positive/TH‐positive fiber plexa were shown. When a recently developed direct triple‐labelling procedure with biotinconjugated mouse secondary antibodies in conjunction with diethylaminocoumarin (DAMC)‐conjugated avidin was employed, presence of GAD/GAL/NT‐ as well as GAD/GRF/NT‐containing varicosities could be demonstrated close to hypophysial portal vessels. In colchicine‐pretreated animals, GAD was shown to coexist with TH, NT, or GAL in cell bodies in both the dorsomedial and ventrolateral domains of the arcuate nucleus, but with GRF only in the ventrolateral division. ChAT‐positive neurons in the ventrolateral region were also TH‐positive. In the ventrolateral arcuate nucleus, triple‐labelling followed by elution‐restaining showed GAD/NT/GAL/TH‐immunoreactivities in the same cells. Similarly, double‐labelling with two following elution‐restaining steps showed several NT/GAL/GRF/TH‐containing cell bodies in this part of the arcuate nucleus. GAD‐positive cells in the anterior hypothalamic periventricular area and fibers in the pituitary neurointermediate lobe were also TH‐positive. The results demonstrate complex patterns of storage of chemical messengers in neurons of the arcuate nucleus‐median eminence complex. Possible neuroendocrine interactions of these systems in the control of prolactin and growth hormone secretion are discussed.