Chen Wenbo, Zhang Zhen, Dong Haiyan, Jiang Xiaoxue
Department of Biology, Institute of Resources and Environment, Henan Polytechnic University, Jiaozuo, Henan 454000, China.
Department of Biology, Institute of Resources and Environment, Henan Polytechnic University, Jiaozuo, Henan 454000, China.
Comp Biochem Physiol B Biochem Mol Biol. 2015 Jun;184:1-9. doi: 10.1016/j.cbpb.2015.01.005. Epub 2015 Feb 7.
Selenoprotein W (SelW) is a low molecular weight and selenocysteine containing protein with redox activity involved in the antioxidant response. In the present study, the full-length cDNA of goldfish (Carassius auratus) selenoprotein W (gfSelW) was successfully cloned from the liver tissue by rapid amplification of cDNA ends technique. The obtained gfSelW cDNA was 730 bp long with a 79 bp 5'-untranslated region (UTR), a 390 bp 3'-UTR containing the consensus polyadenylation signal AATAAA and a 261 bp open reading frame coding a protein of 86 amino acid residues. gfSelW mRNA was observed in all regions of brain and peripheral tissues by semi-quantitative RT-PCR, and the most abundant was detected in testis. After fasting for 1 week, gfSelW mRNA expression levels were significantly decreased compared to the fed group in hypothalamus and liver. After refeeding for 7 days, gfSelW mRNA expression levels were increased back. Furthermore, the mRNA expressions of gfSelW in hypothalamus and liver were varied in periprandial changes and significantly up-regulated after meal 2 h and 4 h, respectively. With cadmium exposure for 24 h, gfSelW mRNA expression levels in gill and leucocytes were significantly decreased at different cadmium concentrations changing from 0.5 ppm to 10 ppm. However, the gfSelW mRNA expression level was sharply increased in liver, relatively to the control about 4.98-fold at 0.5 ppm. The results in this study provide molecular characterization of SelW in goldfish and imply that SelW mRNA expression may be associated with metabolic status and oxidative stress and regulated by metabolic factors and cadmium in fish.
硒蛋白W(SelW)是一种低分子量且含硒代半胱氨酸的蛋白质,具有参与抗氧化反应的氧化还原活性。在本研究中,通过cDNA末端快速扩增技术成功地从金鱼(Carassius auratus)肝脏组织中克隆出硒蛋白W(gfSelW)的全长cDNA。所获得的gfSelW cDNA长730 bp,具有79 bp的5'非翻译区(UTR)、一个390 bp的3'UTR,其中包含共有多聚腺苷酸化信号AATAAA以及一个261 bp的开放阅读框,编码一个含86个氨基酸残基的蛋白质。通过半定量RT-PCR在脑和外周组织的所有区域均观察到gfSelW mRNA,其中在睾丸中检测到的含量最为丰富。禁食1周后,与喂食组相比,下丘脑和肝脏中gfSelW mRNA表达水平显著降低。再喂食7天后,gfSelW mRNA表达水平回升。此外,下丘脑和肝脏中gfSelW的mRNA表达在围餐期变化中有所不同,分别在进食后2小时和4小时显著上调。镉暴露24小时后,鳃和白细胞中gfSelW mRNA表达水平在0.5 ppm至10 ppm的不同镉浓度下均显著降低。然而,在肝脏中,gfSelW mRNA表达水平急剧增加,在0.5 ppm时相对于对照组约增加4.98倍。本研究结果提供了金鱼中SelW的分子特征,并表明SelW mRNA表达可能与代谢状态和氧化应激相关,且受鱼类代谢因子和镉的调控。