Gonadotropin-releasing hormone (GnRH) is a conservative neurodecapeptide fam-ily, which plays a crucial role in regulating the gonad development and in controlling the final sexual maturation in vertebrate. Two differ...Gonadotropin-releasing hormone (GnRH) is a conservative neurodecapeptide fam-ily, which plays a crucial role in regulating the gonad development and in controlling the final sexual maturation in vertebrate. Two differing cGnRH-II cDNAs of common carp, namely cGnRH-II cDNA1 and cDNA2, were firstly cloned from the brain by rapid amplification of cDNA end (RACE) and reverse transcription- polymerase chain reaction (RT-PCR). The length of cGnRH-II cDNA1 and cDNA2 was 622 and 578 base pairs (bp), respectively. The cGnRH-II pre-cursors encoded by two cDNAs consisted of 86 amino acids, including a signal peptide, cGnRH-II decapeptide and a GnRH-associated peptide (GAP) linked by a Gly-Lys-Arg proteolytic site. The results of intron trapping and Southern blot showed that two differing cGnRH-II genes in common carp genome were further identified, and that two genes might exist as a single copy. The multi-gene coding of common carp cGnRH-II gene offered novel evidence for gene duplica-tion hypothesis. Using semi-quantitative RT-PCR, expression and relative expression levels of cGnRH-II genes were detected in five dissected brain regions, pituitary and gonad of common carp. With the exception of no mRNA2 in ovary, two cGnRH-II genes could be expressed in all the detected tissues. However, expression levels showed an apparent difference in different brain regions, pituitary and gonad. According to the expression characterization of cGnRH-II genes in brain areas, it was presumed that cGnRH-II might mainly work as the neurotransmitter and neuromodulator and also operate in the regulation for the GnRH releasing. Then, the ex-pression of cGnRH-II genes in pituitary and gonad suggested that cGnRH-II might act as the autocrine or paracrine regulator.展开更多
基金This work was supported by the'863'High Technology Project of China(Grant No.2001AA213101,2001AA212281)the National Natural Science Foundation of China(Grant No.200102006)+1 种基金the'973'Project of the Ministry of ScienceTechnology(Grant No.2001 CB 109006).
文摘Gonadotropin-releasing hormone (GnRH) is a conservative neurodecapeptide fam-ily, which plays a crucial role in regulating the gonad development and in controlling the final sexual maturation in vertebrate. Two differing cGnRH-II cDNAs of common carp, namely cGnRH-II cDNA1 and cDNA2, were firstly cloned from the brain by rapid amplification of cDNA end (RACE) and reverse transcription- polymerase chain reaction (RT-PCR). The length of cGnRH-II cDNA1 and cDNA2 was 622 and 578 base pairs (bp), respectively. The cGnRH-II pre-cursors encoded by two cDNAs consisted of 86 amino acids, including a signal peptide, cGnRH-II decapeptide and a GnRH-associated peptide (GAP) linked by a Gly-Lys-Arg proteolytic site. The results of intron trapping and Southern blot showed that two differing cGnRH-II genes in common carp genome were further identified, and that two genes might exist as a single copy. The multi-gene coding of common carp cGnRH-II gene offered novel evidence for gene duplica-tion hypothesis. Using semi-quantitative RT-PCR, expression and relative expression levels of cGnRH-II genes were detected in five dissected brain regions, pituitary and gonad of common carp. With the exception of no mRNA2 in ovary, two cGnRH-II genes could be expressed in all the detected tissues. However, expression levels showed an apparent difference in different brain regions, pituitary and gonad. According to the expression characterization of cGnRH-II genes in brain areas, it was presumed that cGnRH-II might mainly work as the neurotransmitter and neuromodulator and also operate in the regulation for the GnRH releasing. Then, the ex-pression of cGnRH-II genes in pituitary and gonad suggested that cGnRH-II might act as the autocrine or paracrine regulator.
基金This work was supported by the National Marine 863 Projects of China (Nos. 2003AA621010 2003AA621110) the NationalNatural Science Foundation of China (No. 39970586) Key Project of Scientific and Technological Research of the Ministry ofEducation (No
文摘研究斜带石斑鱼生长激素分泌及其 mRNA 表达的调控规律对于性别分化的控制、临床药物的选择,以及石斑鱼的增养殖等均具有重要的理论意义和实践意义。本文应用静态孵育系统, 采用放射免疫测定法和化学发光液相杂交实验, 研究GnRH 和 DA 对斜带石斑鱼 GH 分泌、GH mRNA 合成的调控作用。100 nmol/L sGnRH 作用斜带石斑鱼脑垂体碎片 1~24 h,明显促进GH的释放和GH mRNA的合成, 并具有时间依存性; 10 nmol/L^1 μmol/L sGnRH作用1 h 能明显促进斜带石斑鱼脑垂体释放GH,促进GH mRNA的合成, 表现出明显的剂量效应。100 nmol/L、1 μmol/L mGnRH 作用1 h 以一定的剂量依存方式促进 GH 的释放、促进 GH mRNA的合成,但 mGnRH 的效应比相应剂量的 sGnRH 的作用弱。APO 为 DA 受体的非选择性激动剂,不同剂量APO 对斜带石斑鱼脑垂体碎片的作用结果显示, 10 nmol/L^1 μmol/L APO 以剂量依存方式促进斜带石斑鱼脑垂体碎片释放GH、促进 GH mRNA的合成;1 μmol/L APO 作用 12 h 以上明显促进GH 的释放和 GH mRNA的合成,并随时间的延长而增加。与 sGnRH 对斜带石斑鱼 GH 释放、GH mRNA合成的作用相比, APO 的作用较弱。本文研究结果证实GnRH 和 DA能促进斜带石斑鱼脑垂体GH 释放和GH mRNA合成。
