Immunoreactive LH-RH neurons in the hypothalamus identified by light and fluorescent microscopy

The sites of luteinizing hormone-releasing hormone (LH-RH) neurons and their axonal pathways in the hypothalmi of rats and mice were studied by the immunoglobulin-peroxidase bridge technique and b) the immunofluorescent isothiocyanate technique, using rabbit antiserum to synthetic LH-RH. Both of these techniques give similar results concerning LH-RH localization both in rats and mice. The immunoreactive LH-RH positive neurons are about 18-26 mu in diameter, mostly angular, pyriform or spindle shaped and very rarely oval or round. Most of these neurons are generally scattered in the hypothalamus. In the preoptic and medial preshiasmatic areas, ventromedial, arcuate nuclei and ventrolateral-premammillary body, the neurons are numerous and mostly in groups of 3-6. In other areas of the hypothalamus surrounding the suprachiasmatic area, supraoptic area, paraventricular nuclei and the anterior hypothalamic area, the LH-RH neurons are few and scattered. Very few neurons occur in the median eminence or in the infundibular stem. Major projections of LH-RH neurons apparently originate from preoptic, prechiasmatic and surrounding suprachiasmatic area, and from the anterior hypothalamic area. The fibres run in the fabrillar zone, descend in the external infundibular zone, and finally terminate for the most part around the capillaries of the primary portal plexus of the hypophysis. Other major groups of fibres apparently originate from the arcuate and ventromedial neurons. These fibres bend above the fibrous zone for some distance, ultimately descending in the zona palisadica and zona granulosa of the median eminence, towards the capillaries. The fibers of the ventrolateral-premammillary neurons are seen on the floor of the premammillary recess of 3rd ventricle and apparently terminate close to the capillaries in the infundibular region. Most of the LH-RH nerve fibres terminate near the primary portal plexus of the hypophysis, while some terminate in the pars tuberalis and a few seem to terminate in the 3rd ventricle. The LH-RH antiserum is ineffective after absorption with the antigen, so the staining reaction appears to be specific for LH-RH.