Localization of immunoreactive lamprey gonadotropin-releasing hormone in the rat brain☆

Abstract

A highly specific antiserum against lamprey gonadotropin-releasing hormone (GnRH) was used to localize l-GnRH in areas of the rat brain associated with reproductive function. Immunoreactive l-GnRH-like neurons were observed in the ventromedial preoptic area (POA), the region of the diagonal band of Broca and the organum vasculosum lamina terminalis, with fiber projections to the rostral wall of the third ventricle and the organum vasculosum lamina terminalis. Another population of l-GnRH-like neurons was localized in the dorsomedial and lateral POA, with nerve fibers projecting caudally and ventrally to terminate in the external layer of the median eminence. Other fibers apparently projected caudally and circumventrically to terminate around the cerebral aqueduct in the mid-brain central gray. By using a highly specific antiserum directed against mammalian luteinizing hormone-releasing hormone (m-LHRH), the localization of the LHRH neuronal system was compared to that of the l-GnRH system. There were no LHRH neurons in the dorsomedial or the lateral region of the POA that contained the l-GnRH neurons. As expected, there was a large population of LHRH neurons in the ventromedial POA associated with the diagonal band of Broca and organum vasculosum lamina terminalis. In both of these regions, there were many more LHRH neurons than l-GnRH neurons and the LHRH neurons extended more dorsally and laterally than the l-GnRH neurons. The LHRH neurons seemed to project to the median eminence in the same areas as those that were innervated by the l-GnRH neurons. Absorption studies indicated that l-GnRH cell bodies were eliminated by adding 1 μg of either l-GnRH-I or 1-GnRH-III, but not m-LHRH to the antiserum before use. Fibers were largely eliminated by the addition of 1 μg l-GnRH-III to the antiserum. No chicken GnRH-II neurons or nerve fibers could be visualized by immunostaining. Because the antiserum recognized GnRH-I and GnRH-III equally, we have visualized an l-GnRH system in rat brain. The results are consistent with the presence of either one or both of these peptides within the rat hypothalamus. Because l-GnRH-I has only weak nonselective gonadotropin-releasing activity, whereas l-GnRH-III is a highly selective releaser of follicle-stimulating hormone, and because l-GnRH neurons are located in areas known to control follicle-stimulating hormone release selectively, our results support the hypothesis that l-GnRH-III, or a closely related peptide, may be mammalian follicle-stimulating hormone-releasing factor.