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陈文波, 王鑫, 周雅璐, 董海燕, 林浩然, 李文笙. 2011: 尼罗罗非鱼Orexin前体基因的克隆、组织分布及其在摄食调控中的表达. 动物学研究, 32(3): 285-292. DOI: 10.3724/SP.J.1141.2011.03285
引用本文: 陈文波, 王鑫, 周雅璐, 董海燕, 林浩然, 李文笙. 2011: 尼罗罗非鱼Orexin前体基因的克隆、组织分布及其在摄食调控中的表达. 动物学研究, 32(3): 285-292. DOI: 10.3724/SP.J.1141.2011.03285
CHEN Wen-Bo, WANG Xin, ZHOU Ya-Lu, DONG Hai-Yan, LIN Hao-Ran, LI Wen-Sheng. 2011. Molecular cloning, tissue distribution and the expression in the regulation of food intake of prepro-orexin in Nile tilapia (Oreochromis niloticus). Zoological Research, 32(3): 285-292. DOI: 10.3724/SP.J.1141.2011.03285
Citation: CHEN Wen-Bo, WANG Xin, ZHOU Ya-Lu, DONG Hai-Yan, LIN Hao-Ran, LI Wen-Sheng. 2011. Molecular cloning, tissue distribution and the expression in the regulation of food intake of prepro-orexin in Nile tilapia (Oreochromis niloticus). Zoological Research, 32(3): 285-292. DOI: 10.3724/SP.J.1141.2011.03285

尼罗罗非鱼Orexin前体基因的克隆、组织分布及其在摄食调控中的表达

Molecular cloning, tissue distribution and the expression in the regulation of food intake of prepro-orexin in Nile tilapia (Oreochromis niloticus)

  • 摘要: 该文采用RT-PCR和cDNA末端快速扩增技术(rapid-amplification of cDNA ends, RACE)的方法, 从尼罗罗非鱼(Oreochromis niloticus)下丘脑总RNA中获得了尼罗罗非鱼Orexin前体基因的cDNA全长序列。该 cDNA全长648 bp, 其中开放阅读框的长423 bp,编码Orexin前体蛋白为140个氨基酸, 包括37个氨基酸的信号肽、43个氨基酸的Orexin-A、28个氨基酸的Orexin-B和末尾32个氨基酸组成的功能不详的多肽。采用Real-time PCR 技术对尼罗罗非鱼Orexin前体基因的组织表达模式以及在摄食前后、饥饿和再投喂状态下的表达量变化进行了研究。结果显示, 在脑部和外周等18个组织中都检测到了Orexin前体基因的表达, 其中在下丘脑中表达量最高; 在摄食前后, 尼罗罗非鱼Orexin前体基因的表达量显著低于在摄食状态中; 饥饿2、4、6和8 d后, Orexin前体基因在下丘脑中的表达量与正常投喂组相比均显著升高, 饥饿4 d再投喂后,表达量又恢复至正常水平。这些结果表明, Orexin在尼罗罗非鱼摄食中可能有着重要的调节作用。

     

    Abstract: We cloned the full length of tilapia prepro-orexin cDNA using RT-PCR and rapid-amplification of cDNA ends (RACE). The full-length of prepro-orexin cDNA was 648 bp containing an open reading frame of 423 bp. The 140 amino acid prepro-orexin protein included a 37 AA signal peptide, a 43 AA Orexin-A and, and 28 AA Orexin-B and the end of the 32 AA peptide of unknown function. The expression of prepro-orexin on tissue distribution, peri-prandial changes, starvation and re-feeding were quantified by real-time PCR. We found that prepro-orexin mRNA was present in all tissues tested and that the highest level was observed in hypothalamus. Expression levels were significantly higher at mealtime (0 h) than before (−2 h, −1 h) and after (+1 h, +2 h) mealtime. Fasting for 2, 4, 6 and 8 d caused significant increases in prepro-orexin mRNA expression in the hypothalamus, and after re-feeding, expression levels of prepro-o rexin mRNA returned to the same level compared to that in the fed group.

     

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