Identification of His5,Trp7,Tyr8-GnRH (chicken GnRH II) in amphibian brain |
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| Authors: | J A King R P Millar |
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| Affiliation: | 1. M.R.C. Regulatory Peptides Research Unit, Department of Chemical Pathology University of Cape Town Medical School Observatory 7925, Cape Town, South Africa;2. Groote Schuur Hospital Observatory 7925, Cape Town, South Africa;1. Evolution des Régulations Endocriniennes, UMR 7221 CNRS, Muséum National d’Histoire Naturelle, Sorbonne Université, Paris, France;2. Institut de systématique et Evolution, UMR 7205 CNRS, UMPC, EPHE, Muséum National d’Histoire Naturelle, Sorbonne Université, Paris, France;4. Department of Cell Biology and Ecology, CIBUS, Faculty of Biology, University of Santiago de Compostela, Spain;5. Biologie Intégrative des Organismes Marins, UMR 7232 CNRS, Observatoire Océanologique, Université Pierre et Marie Curie, Sorbonne Université, Banyuls-sur-Mer, France;6. Laboratory of Molecular Mechanisms of Neuronal Interactions, Sechenov Insitute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St. Petersburg, Russia;1. Graduate School of Natural Science and Technology, Okayama University, 3-1-1 Tsushima-naka, Okayama 700-8530, Japan;2. Department of Biology, Faculty of Science, Okayama University, 3-1-1 Tsushima-naka, Okayama 700-8530, Japan;3. Graduate School of Integrated Sciences for Life, Hiroshima University, 1-3-2 Kagamiyama, Higashi-Hiroshima City, Hiroshima 739-8511, Japan;4. Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Aoba-Ku, Sendai 980-8577, Japan |
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| Abstract: | GnRH immunoreactive and bioactive peptides in Xenopus laevis brain extract were investigated by high performance liquid chromatography (HPLC), radioimmunoassay with region-specific antisera raised against GnRH (mammalian), His5,Trp7,Tyr8-GnRH (chicken II) and Tyr3,Leu5,Glu6,Trp7,Lys8-GnRH (lamprey), and by assessment of biological activity. Two immunoreactive peptides eluted in the same positions as GnRH and His5,Trp7,Tyr8-GnRH respectively in HPLC systems which were specifically designed to separate four known natural vertebrate GnRHs (mammalian, chicken I and II, salmon). The immunological properties of these two immunoreactive peaks, determined by relative interaction with three region-specific antisera raised against mammalian GnRH and two specific His5,Trp7,Tyr8-GnRH antisera, were identical to those of GnRH and His5,Trp7,Tyr8-GnRH. The immunoreactive peak co-eluting with His5,Trp7,Tyr8-GnRH represented approximately one-third of the total brain GnRH. Both immunoreactive peaks stimulated luteinizing hormone (LH) release in a chicken dispersed pituitary cell bioassay, and the amounts of LH release stimulated by the two peaks were appropriate for these peaks being GnRH and His5,Trp7,Tyr8-GnRH. A small hydrophobic peak with GnRH immunoreactivity eluted in the same position as Trp7,Leu8-GnRH (salmon), while Gln8-GnRH (chicken I) and lamprey GnRH were not detected. Two additional rather hydrophilic peptides cross-reacted with a COOH-terminus-directed antiserum and had LH-releasing activity. LH-releasing activity was also detected in hydrophobic HPLC fractions. In summary, these data provide evidence for the presence of both GnRH and a second peptide with properties identical to His5,Trp7,Tyr8-GnRH in X. laevis brain.(ABSTRACT TRUNCATED AT 250 WORDS) |
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