Ashihara M, Suzuki M, Kubokawa K, Yoshiura Y, Kobayashi M, Urano A, Aida K
Department of Fisheries, Faculty of Agriculture, University of Tokyo, Japan.
J Mol Endocrinol. 1995 Aug;15(1):1-9. doi: 10.1677/jme.0.0150001.
Salmon gonadotropin-releasing hormone (sGnRH) is considered to have an important role in the control of reproduction in salmonid fish. As a basis for understanding the physiological functioning of sGnRH at the molecular level, we characterized the nucleotide sequences of two types of cDNAs encoding the precursors of sGnRH in sockeye salmon (ss), Oncorhynchus nerka, by a cloning strategy based on reverse transcription-PCR. The two types of cDNAs are referred to as ss-pro-sGnRH-I and -II, and consisted of 435 and 481 bases respectively. Both precursors are predicted to contain a signal peptide, the hormone and a GnRH-associated peptide that is attached to the hormone via a Gly-Lys-Arg sequence. The presence of two types of mRNAs hybridizing with either cDNA was confirmed by Northern blot analysis of brain RNA from sockeye salmon, masu salmon, O. masou, and rainbow trout, O. mykiss. The ss-pro-sGnRH-I cDNA had 97.2% and 82.8% overall identity with sGnRH cDNA from masu salmon and putative sGnRH cDNA deduced from the gene of the Atlantic salmon, Salmo salar respectively, whereas the ss-pro-sGnRH-II cDNA had 80.0% and 91.2% overall identity with the former and the latter respectively. The nucleotide sequences of ss-sGnRH-I and -II cDNAs showed less similarity (79.3%). These results indicated that each salmonid species possesses two differing sGnRH genes. The results of Southern blot analysis using genomic DNA extracted from individuals support this evidence in sockeye salmon, masu salmon and rainbow trout.
鲑鱼促性腺激素释放激素(sGnRH)被认为在鲑科鱼类的繁殖控制中发挥着重要作用。作为在分子水平上理解sGnRH生理功能的基础,我们通过基于逆转录聚合酶链反应的克隆策略,对红大麻哈鱼(Oncorhynchus nerka)中编码sGnRH前体的两种cDNA的核苷酸序列进行了表征。这两种cDNA分别被称为ss-pro-sGnRH-I和-II,分别由435和481个碱基组成。两种前体均预计包含一个信号肽、该激素以及一个通过甘氨酸-赖氨酸-精氨酸序列与该激素相连的GnRH相关肽。通过对红大麻哈鱼、马苏大麻哈鱼(O. masou)和虹鳟(O. mykiss)脑RNA的Northern印迹分析,证实了存在与任一cDNA杂交的两种mRNA。ss-pro-sGnRH-I cDNA与马苏大麻哈鱼的sGnRH cDNA以及从大西洋鲑(Salmo salar)基因推导的推定sGnRH cDNA的总体一致性分别为97.2%和82.8%,而ss-pro-sGnRH-II cDNA与前者和后者的总体一致性分别为80.0%和91.2%。ss-sGnRH-I和-II cDNA的核苷酸序列显示出较低的相似性(79.3%)。这些结果表明,每个鲑科物种都拥有两个不同的sGnRH基因。使用从个体中提取的基因组DNA进行的Southern印迹分析结果在红大麻哈鱼、马苏大麻哈鱼和虹鳟中支持了这一证据。