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黄颡鱼中过氧化物酶体增殖物激活受体γ(PPARγ):分子特征、mRNA表达及体内外胰岛素对其转录调控

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

作者信息

Zheng Jia-Lang, Zhuo Mei-Qin, Luo Zhi, Pan Ya-Xiong, Song Yu-Feng, Huang Chao, Zhu Qing-Ling, 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

Gen Comp Endocrinol. 2015 Feb 1;212:51-62. doi: 10.1016/j.ygcen.2014.12.020. Epub 2015 Jan 28.

Abstract

Peroxisome proliferator-activated receptor gamma (PPARγ) is ligand-inducible transcription factor and has important roles in lipid metabolism, cell proliferation and inflammation. In the present study, yellow catfish Pelteobagrus fulvidraco PPARγ cDNA was isolated from liver by RT-PCR and RACE, and its molecular characterization and transcriptional regulation by insulin in vivo and in vitro were determined. The generation of PPARγ1 and PPARγ2 was due to alternative promoter of PPARγ gene. PPARγ1 and PPARγ2 mRNA covered 2426 bp and 2537 bp, respectively, with an open reading frame (ORF) of 1584 bp encoding 527 amino acid residues. Yellow catfish PPARγ gene was organized in a manner similar to that of their mammalian homologs, implying a modular organization of the protein's domains. A comparison between the yellow catfish PPARγ amino acid sequence and the correspondent sequences of several other species revealed the identity of 55-76.2%. Two PPARγ transcripts (PPARγ1 and PPARγ2) mRNAs were expressed in a wide range of tissues, but the abundance of each PPARγ mRNA showed the tissue- and developmental stage-dependent expression patterns. Intraperitoneal injection of insulin in vivo significantly stimulated the mRNA expression of total PPARγ and PPARγ1, but not PPARγ2 in the liver of yellow catfish. In contrast, incubation of hepatocytes with insulin in vitro increased the mRNA levels of PPARγ1, PPARγ2 and total PPARγ. To our knowledge, for the first time, the present study provides evidence that PPARγ1 and PPARγ2 are differentially expressed with and among tissues during different developmental stages and also regulated by insulin both in vivo and in vitro, which serves to increase our understanding on PPARγ physiological function in fish.

摘要

过氧化物酶体增殖物激活受体γ(PPARγ)是一种配体诱导型转录因子,在脂质代谢、细胞增殖和炎症中发挥重要作用。在本研究中,通过RT-PCR和RACE从黄颡鱼(Pelteobagrus fulvidraco)肝脏中分离出PPARγ cDNA,并确定了其分子特征以及胰岛素在体内和体外对其转录的调控。PPARγ1和PPARγ2的产生是由于PPARγ基因的可变启动子。PPARγ1和PPARγ2的mRNA分别覆盖2426 bp和2537 bp,开放阅读框(ORF)为1584 bp,编码527个氨基酸残基。黄颡鱼PPARγ基因的组织方式与其哺乳动物同源物相似,这意味着该蛋白质结构域具有模块化组织。黄颡鱼PPARγ氨基酸序列与其他几个物种相应序列的比较显示,同一性为55 - 76.2%。两种PPARγ转录本(PPARγ1和PPARγ2)的mRNA在多种组织中表达,但每种PPARγ mRNA的丰度呈现出组织和发育阶段依赖性的表达模式。体内腹腔注射胰岛素显著刺激了黄颡鱼肝脏中总PPARγ和PPARγ1的mRNA表达,但对PPARγ2没有影响。相反,体外将肝细胞与胰岛素孵育可增加PPARγ1、PPARγ2和总PPARγ的mRNA水平。据我们所知,本研究首次提供证据表明,PPARγ1和PPARγ2在不同发育阶段的组织内和组织间存在差异表达,并且在体内和体外均受胰岛素调控,这有助于增进我们对鱼类中PPARγ生理功能的理解。

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