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The origins of the dopamine (DA)-containing nerve terminals in the external layer of the median eminence and in the neurointermediate pituitary were determined in rats by a combination of retrograde labeling with wheat germ agglutinin (WGA) and immunohistochemistry for tyrosine hydroxylase (TH). Biotinylated WGA (b-WGA) was injected into the posterior pituitary (group 1) and into the median eminence (group 2). In group 1 animals, all the magnocellular neurons of the paraventricular, supraoptic, and accessory nuclei, and many parvicellular neurons in the rostral periventricular region (RPR) were labeled with WGA. In group 2 animals, many neurons were labeled in the arcuate nucleus and the RPR, and in a small population of the preoptic-septal region. In group 1 animals, about 39% of TH neurons in the RPR were labeled with WGA, whereas only a few TH neurons (1%) in the arcuate nucleus were labeled with WGA. In group 2 animals, on the contrary, almost all TH neurons (73%) in the arcuate nucleus carried WGA, whereas in the RPR, only some of the TH neurons (19%) were labeled with WGA. It is concluded that DA neurons involved in the hypothalamic-anterior pituitary axis are located in the arcuate nucleus; those involved in neuro-intermediate lobe function in the RPR.

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LHRH neurons and fiber tracts have been studied immunohistochemically in rats. After the intraventricular infusion of colchicine, a number of immunoreactive LHRH neurons were consistently evident in the preoptico-septal area and in the diagonal band of Broca. The immunoreactive fiber tracts originating in those areas were classified into two groups: preoptico-terminal and preoptico-infundibular tracts. The former terminated in the OVLT. The latter was further divided into the mediobasal and laterobasal tracts; both were found to terminate in the median eminence. The mediobasal tract ran posteriorly through the ventral portion of the lateral periventricular area. The laterobasal tract ran posteriorly in the medial forebrain bundle and was accompanied by several immunoreactive neurons. The immunoreactive cell bodies were further scattered in the base of the tuberal hypothalamic area. An anterior bilateral Halász cut in the retrochiasmatic region was followed by a great reduction of the immunoreactive materials in the median eminence and by the appearance of the immunoreactive perikarya in the preoptico-septal area and in the diagonal band of Broca. A horizontal hypothalamic cut, on the other hand, which disconnected the tubero-infundibular tract between the median eminence and the arcuate nuclei, caused neither reduction of the immunoreactive materials in the median eminence nor appearance of the immunoreactive perikarya in the arcuate nuclei. Those results indicate that the bulk of the immunoreactive LHRH is synthesized in the anterior hypothalamic area and transported by the preoptico-terminal and -infundibular tracts but a minor part is produced in the cell bodies located in the basal tuberal hypothalamus and transported to the median eminence.

The morphological substrate for the central mechanisms that control growth hormone (GH) release in the rat hypothalamus was investigated immunohistochemically by light and electron microscopy. In electron-microscopic studies, a dual immunolabeling technique was employed to demonstrate pairs of peptides, i.e. rat hypothalamic growth hormone-releasing factor (rhGRF) and somatostatin (SRIH), rhGRF and substance P (SP), and rhGRF and methionine-enkephalin-Arg⁶-Gly⁷-Leu⁸ (Enk-8), in different neuronal structures. Immunoreactivity of rhGRF was detected as silver-gold particles and those of the other substances as diaminobenzidine products by preembedding immunostaining procedures. In the external layer of the median eminence, axonal terminals immunolabeled for rhGRF and for SRIH showed the same pattern of distribution and close proximity. The neuronal inputs to GRF cell bodies in the arcuate nucleus were examined, and SRIH, SP and Enk-8 fibers with varicosities were found to form dense networks around the perikarya of GRF neurons, suggesting the presence of synaptic associations. Axonal terminals immunolabeled for SRIH, SP or Enk-8, and unlabeled terminals appeared to form coincidental synaptic junctions on GRF perikarya. These findings suggest that the central regulation of GH release occurs at the levels of the median eminence and the cell bodies.