Identification, tissue distribution and regulation of preproghrelin in the brain and gut of Schizothorax prenanti

Regulatory Peptides Pub Date : 2013-09-10 DOI:10.1016/j.regpep.2013.07.002

RongBin Wei, Tao Liu, ChaoWei Zhou, XingDong Zhang, DengYue Yuan, Tao Wang, FangJun Lin, Hu Chen, HongWei Wu, ZhiQiong Li

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引用次数: 19

Abstract

Ghrelin is an important gastrointestinal hormone involved in the regulation of feeding in both mammals and fish. In this study, the preproghrelin cDNA sequence was cloning in the gut of Schizothorax prenanti (S. prenanti). The preproghrelin gene, encoding 103-amino acids, was strongly expressed in the gut and brain using real-time quantitative RT-PCR (qPCR). The S. prenanti preproghrelin was detected in embryonic developmental stages. Further, it was detectable in unfertilized eggs, suggesting that ghrelin could be classified as maternal mRNA. An experiment was conducted to determine the expression profile of ghrelin during post-feeding and fasting status of the brain and gut. The results revealed a significant postprandial decrease in ghrelin mRNA expression in the gut 6 h post-feeding (hpf) and brain (1.5 and 9 hpf) compared to an unfed control group, indicating that food intake and processing affect the regulation of expression of ghrelin in S. prenanti. The constructed recombinant plasmid pMD-19 T-ghrelin was transformed to Escherichia coli BL21 and induced with IPTG, and the expressed product was identified by SDS-PAGE. The prokaryotic expression vector for ghrelin was constructed successfully, and fusion protein was expressed in E. coli BL21, which laid the foundation for the further study on the function of this protein and its mechanism. Overall, our results provide evidence for a highly conserved structure and biological actions of ghrelin in S. prenanti. Further studies are required to identify the tissue specific functions of ghrelin in S. prenanti.

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裂腹鱼脑、肠前促生长素的鉴定、组织分布及调控

胃饥饿素是一种重要的胃肠激素，参与哺乳动物和鱼类的摄食调节。本研究在南裂胸(Schizothorax prenanti, S. prenanti)肠道中克隆了前促生长素cDNA序列。利用实时定量RT-PCR (qPCR)技术，在肠道和大脑中强烈表达了编码103个氨基酸的preproghrelin基因。在胚胎发育阶段检测到S. prenanti preproghrelin。此外，在未受精卵中可检测到ghrelin，这表明ghrelin可以归类为母体mRNA。本实验旨在确定胃饥饿素在饲喂后和禁食状态下大脑和肠道的表达谱。结果显示，与未喂食的对照组相比，摄食后6小时肠道(hpf)和大脑(1.5和9 hpf)胃饥饿素mRNA的餐后表达显著降低，表明食物摄入和加工影响了胃饥饿素表达的调节。将构建的重组质粒pMD-19转化到大肠杆菌BL21中，经IPTG诱导，表达产物经SDS-PAGE鉴定。成功构建了ghrelin原核表达载体，并在大肠杆菌BL21中表达了融合蛋白，为进一步研究该蛋白的功能及其作用机制奠定了基础。综上所述，我们的研究结果为胃饥饿素的高度保守结构和生物学作用提供了证据。ghrelin在S. prenanti中的组织特异性功能有待进一步研究。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Regulatory Peptides 医学-内分泌学与代谢

自引率

0.00%

发文量

审稿时长

2 months

期刊介绍： Regulatory Peptides provides a medium for the rapid publication of interdisciplinary studies on the physiology and pathology of peptides of the gut, endocrine and nervous systems which regulate cell or tissue function. Articles emphasizing these objectives may be based on either fundamental or clinical observations obtained through the disciplines of morphology, cytochemistry, biochemistry, physiology, pathology, pharmacology or psychology.