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黄颡鱼中过氧化物酶体增殖物激活受体α1:分子特征、mRNA组织表达及胰岛素在体内外的转录调控

Peroxisome proliferator-activated receptor alpha1 in yellow catfish Pelteobagrus fulvidraco: molecular characterization, mRNA tissue expression and transcriptional regulation by insulin in vivo and in vitro.

作者信息

Zheng Jia-Lang, Zhuo Mei-Qin, Luo Zhi, Song Yu-Feng, Pan Ya-Xiong, Huang Chao, Hu Wei, Chen Qi-Liang

机构信息

National Engineering Research Center of Marine Facilities Aquaculture, Zhejiang Ocean University, Zhoushan 316022, China; Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture of P.R.C., Fishery College, Huazhong Agricultural University, Wuhan 430070, China; Freshwater Aquaculture Collaborative Innovative Centre of Hubei Province, Wuhan 430070, China.

Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture of P.R.C., Fishery College, Huazhong Agricultural University, Wuhan 430070, China; Freshwater Aquaculture Collaborative Innovative Centre of Hubei Province, Wuhan 430070, China.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2015 May;183:58-66. doi: 10.1016/j.cbpb.2015.01.004. Epub 2015 Jan 30.

Abstract

Peroxisome proliferator-activated receptor alpha1 (PPARα1) cDNA was isolated from liver of yellow catfish Pelteobagrus fulvidraco by RT-PCR and RACE. Its molecular characterization, tissue expression and transcriptional regulation by insulin in vitro and in vivo were determined. PPARα1 mRNA covered 1879 bp, with an open reading frame (ORF) of 1410 bp encoding 469 amino acid residues, a 5'-untranslated region (UTR) of 49 bp, and a 3'-UTR of 421 bp. PPARα1 consisted of 4 domains, the A/B domain, DNA-binding domain (DBD), D domain, and ligand-binding domain (LBD). The predicted tertiary structure of yellow catfish PPARα1 showed an increased size of the main cavity that was made up of side chains from helices 3, 5, 10, 11, and 12. Changes of PPARα1 structure might affect binding of mammalian PPARα1-specific ligand and cofactor in yellow catfish and may endow yellow catfish PPARα1 with new ligand-independent or -dependent transactivation activity. PPARα1 was differentially expressed in various tissues during development. Furthermore, intraperitoneal injection in vivo and incubation in vitro of insulin reduced the mRNA expression of PPARα1 in the liver and hepatocytes of yellow catfish. Based on the observation above, the present study provides evidence that PPARα1 is differentially expressed within and among tissues during three developmental stages and also regulated by insulin both in vivo and in vitro, which warrants further investigation of PPARα1 physiological function in fish.

摘要

通过逆转录聚合酶链反应(RT-PCR)和快速扩增cDNA末端(RACE)技术,从黄颡鱼(Pelteobagrus fulvidraco)肝脏中分离出过氧化物酶体增殖物激活受体α1(PPARα1)的cDNA。测定了其分子特征、组织表达以及在体内外胰岛素对其转录调控的影响。PPARα1的mRNA全长1879 bp,开放阅读框(ORF)为1410 bp,编码469个氨基酸残基,5'-非翻译区(UTR)为49 bp,3'-UTR为421 bp。PPARα1由4个结构域组成,即A/B结构域、DNA结合结构域(DBD)、D结构域和配体结合结构域(LBD)。预测的黄颡鱼PPARα1三级结构显示,由螺旋3、5、10、11和12的侧链组成的主腔尺寸增大。PPARα1结构的变化可能会影响哺乳动物PPARα1特异性配体和辅因子与黄颡鱼PPARα1的结合,并可能赋予黄颡鱼PPARα1新的非配体依赖性或配体依赖性反式激活活性。PPARα1在发育过程中的不同组织中差异表达。此外,体内腹腔注射和体外胰岛素孵育均可降低黄颡鱼肝脏和肝细胞中PPARα1的mRNA表达。基于上述观察结果,本研究提供了证据表明PPARα1在三个发育阶段的组织内和组织间差异表达,并且在体内外均受胰岛素调节,这值得进一步研究PPARα1在鱼类中的生理功能。

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