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首页> 外文期刊>Zoological Science >Steroidogenic acute regulatory protein in eels: cDNA cloning and effects of ACTH and seawater transfer on its mRNA expression
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Steroidogenic acute regulatory protein in eels: cDNA cloning and effects of ACTH and seawater transfer on its mRNA expression

机译:鳗类类固醇激素急性调节蛋白:cDNA的克隆以及ACTH和海水转移对其mRNA表达的影响

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摘要

Steroidogenic acute regulatory protein (StAR) is a key molecule for steroid production by translocating cholesterol from the outer to inner mitochondrial membrane. Two cDNAs of different length encoding StAR was cloned from the head kidney of the eel (Anguilla japonica). In the 3'-untranslated region (UTR) of the longer cDNA, two putative polyadenylation signals were found. The shorter one differed from the longer one solely by the lack of middle of 3'-UTR including the first polyadenylation signal. Reverse transcription-polymerase chain reaction (RT-PCR) that differentiates the two mRNAs showed that the ratio of the two was highly variable among individuals, and no preferential expression was detected between freshwater and seawater eels. The predicted protein consists of 285 amino acid residues with 64-83% identity to other StARs thus far obtained. RT-PCR analyses revealed that eel StAR mRNA was expressed abundantly in the head kidney and gonad, and faintly in the brain; but no expression was detected in the gill, heart, liver, intestine, kidney and skeletal muscle. Plasma cortisol concentration increased, but StAR mRNA content in the head kidney did not change, 3 and 24 h after transfer of freshwater eels to seawater, indicating that the transcriptional regulation of StAR may not be involved in cortisol production after seawater transfer. However, ACTH elevated both plasma cortisol and StAR mRNA levels in the head kidney 1.5 and 4.5 h after injection. Thus, the steroidogenic effect of ACTH is mediated by increased StAR production as observed in mammals.
机译:类固醇生成的急性调节蛋白(StAR)是通过将胆固醇从线粒体外膜向内膜移位而产生类固醇的关键分子。从鳗鱼的头部肾脏克隆了两个不同长度的编码StAR的cDNA。在较长cDNA的3'-非翻译区(UTR)中,发现了两个推定的聚腺苷酸化信号。较短的一个与较长的一个仅由于缺少包括第一聚腺苷酸化信号的3'-UTR的中间而不同。区分这两种mRNA的逆转录聚合酶链反应(RT-PCR)表明,两者的比例在个体之间高度可变,并且在淡水和海水鳗鱼之间未检测到优先表达。预测的蛋白质由285个氨基酸残基组成,与迄​​今获得的其他StAR具有64-83%的同一性。 RT-PCR分析显示,鳗鱼StAR mRNA在头肾和性腺中大量表达,在脑中微弱表达;但在the,心脏,肝脏,肠,肾脏和骨骼肌中未检测到表达。在将淡水鳗鱼转移到海水中后3和24小时,血浆皮质醇浓度增加,但头部肾脏中StAR mRNA的含量没有变化,这表明在海水转移后,StAR的转录调控可能不参与皮质醇的产生。然而,ACTH在注射后1.5和4.5 h升高了头肾的血浆皮质醇和StAR mRNA水平。因此,如在哺乳动物中观察到的,ACTH的类固醇生成作用是通过增加StAR产生的。

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