鳜三种生长抑素cDNA全长克隆、组织表达及其在脑中定位分析
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上海海洋大学农业部淡水水产种质资源重点实验室,上海海洋大学农业部淡水水产种质资源重点实验室,上海海洋大学农业部淡水水产种质资源重点实验室,上海海洋大学农业部淡水水产种质资源重点实验室

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上海市科委基础研究重点项目(09JC1406900);上海市重点学科建设项目(Y1101);上海海洋大学博士启动基金(07274)


Cloning,tissue expression of three somatostatin genes of Siniperca chuatsi and their localizations in brain
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Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture,Shanghai Ocean University,Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture,Shanghai Ocean University,Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture,Shanghai Ocean University,Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture,Shanghai Ocean University

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    摘要:

    利用cDNA末端快速扩增法(RACE)克隆了鳜脑中3种生长抑素前体(preprosomatostatin,PSSⅠ、PSSⅡ和PSSⅢ)cDNA全长序列。结果显示,鳜PSSⅠ cDNA全长647 bp,含开放阅读框369 bp,共编码122个氨基酸,前26个氨基酸为信号肽,C端SS14在脊椎动物中非常保守; PSSⅡ cDNA全长655 bp,开放阅读框387 bp,编码128个氨基酸,前25个氨基酸为信号肽,C端带有[Tyr7,Gly10]SS-14; PSSⅢ cDNA全长823 bp,开放阅读框333 bp,共编码110个氨基酸,前21个氨基酸为信号肽,C端带有[Pro2]SS-14。利用RT-PCR检测了鳜PSS mRNA的组织表达特征,3种PSS mRNA均在脑中表达,PSSⅠ mRNA在其他组织中未检测到表达,PSSⅡ mRNA在食道和胃中大量表达,PSSⅢ mRNA在肾和性腺中大量表达。推测3种PSS除一致参与神经调控外,PSSⅡ还可参与消化作用调控,PSSⅢ参与调控生殖作用。脑组织原位杂交分析表明,PSSⅠ mRNA在下丘脑和延脑中表达,PSSⅡ mRNA在下丘脑中表达,PSSⅢ mRNA在下丘脑、视觉盖、延脑和脊髓中表达。3种PSS均在下丘脑中表达,表明它们可能参与腺垂体激素分泌的调节; 在其他脑区存在表达差异,推测它们还可作为神经递质或调节因子,发挥不同的生物学效应。

    Abstract:

    Somatostatins(SS)play important physiological functions in vertebrate’s growth and development,which belong to a multiple gene family.In this study,the complete cDNA sequences of three distinct preprosomatostatins(PSSⅠ,PSSⅡand PSSⅢ)were isolated from Siniperca chuatsi brain and cloned by means of rapid amplification of cDNA ends(RACE).The full length of PSSⅠ cDNA was 647 bp,which contained 369 bp open reading frame and encoded 122 amino acids with a signal peptide of 26 amino acids.It contained the conservative SS-14 sequence at the Cterminal extremity,which is identical with that of other vertebrates.The full length of PSSⅡ cDNA was 655 bp,which contained 387 bp open reading frame and encoded 128 amino acids with a signal peptide of 25 amino acids.It contained[Tyr7,Gly10]SS-14 at the C-terminal extremity.The full length of PSSⅢ cDNA was 823 bp,which contained 333 bp open reading frame and encoded 110 amino acids with a signal peptide of 21 amino acids.It contained[Pro2]SS-14 at its C-terminal extremity.PSS mRNA expressions in different adult tissues were analyzed by RT-PCR technique.All PSS mRNA were found in brain,while PSSⅠ mRNA wasn’t detected in other peripheral tissues,high levels of PSSⅡ mRNA were also present in esophagus and stomach,PSSⅢ mRNA in kidney and gonad.These suggested that all PSSs participate in neurological regulation in brain,whereas PSSⅡ may also regulate digestion activity,and PSSⅢ regulates reproduction activity.Cellular localization of PSS mRNAs in brain was determined by in situ hybridization,PSSⅠ mRNA was expressed in hypothalamus and medulla oblongata,PSSⅡ mRNA was expressed in hypothalamus,PSSⅢ mRNA was expressed in hypothalamus,optic tectun,medulla oblongata and spinal cord.All PSS mRNA were detected in the hypothalamus,which showed they could be involved in hypothalamic-pituitary regulation.Distinct expression in other brain sections suggested somatostatins may also act as neurotransmitters or hormones,and exhibit different biological effects.

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代威,赵金良,刘俊,薛洋.鳜三种生长抑素cDNA全长克隆、组织表达及其在脑中定位分析[J].水产学报,2012,36(9):1337~1348

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  • 收稿日期:2011-12-20
  • 最后修改日期:2012-02-26
  • 录用日期:2012-08-24
  • 在线发布日期: 2012-09-19
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