DISTRIBUTION OF LUTEINIZING-HORMONE-RELEASING HORMONES-I AND HORMONES-II (LHRH-I AND LHRH-II) IN THE QUAIL AND CHICKEN BRAIN AS DEMONSTRATED WITH ANTIBODIES DIRECTED AGAINST SYNTHETIC PEPTIDES

Polyclonal antibodies were raised in rabbits against polypeptides corresponding to the N-terminal part (heptapeptides) of the two avian gonadotropin-releasing hormones, chicken (c) LHRH-I and -II. These peptides, which were synthesized by the continuous-flow technique, were selected because they contained the smallest number of common amino acid residues. The pGlu-His-Trp-Ser sequence at the C-terminal was suppressed to avoid possible cross-reactions between the antisera. The antisera generated in this way were tested for specificity by solid and liquid phase absorption as well as by antigen spot tests. The antiserum raised against cLHRH-I recognized this peptide preferentially though not exclusively. Some cross-reaction with cLHRH-II was observed in the absorption test, although spotting tests suggested a total specificity. The anti cLHRH-II appeared to be completely specific in all tests. These two antibodies were then used to study the distribution of cLHRH-I and -II immunoreactive structures in the quail and chicken brain. cLHRH-I immunoreactive perikarya were observed in a fairly wide area covering the preoptic-anterior hypothalamic and septal region. By contrast, cLHRH-II cells were confined to a single group located in the dorsal aspects of the occulomotor nuclei, at the junction of the di- and mesencephalon. A sex difference in the number of cLHRH-I cells was detected in the anterior lateral preoptic region of the quail. Fibers immunoreactive for either cLHRH-I or cLHRH-II were widely distributed in the telencephalon, diencephalon, and mesencephalon but showed a specific pattern of anatomical localization. In particular, a high density of cLHRH-I fibers were seen in the external layer of the median eminence, while cLHRH-II fibers were less prominent at this level. Contrary to previous reports, a significant amount of cLHRH-II fibers were however seen throughout the median eminence (mostly external layer). The extensive distribution of both cLHRH-I and -II fibers in the quail and chicken brain is consistent with the potential role played by these peptides in the gonadotropin secretion and in the control of reproductive behavior. The specific role of cLHRH-II remains however elusive at present. (C) 1993 Wiley-Liss, Inc.