文章摘要
张丽晗,罗智,有文静,李丹丹,吴坤,潘亚雄.黄颡鱼FZD家族4个基因的克隆、组织表达及对铜的响应[J].水产学报,2018,42(5):625~632
黄颡鱼FZD家族4个基因的克隆、组织表达及对铜的响应
Molecular characterization and tissue distribution of Frizzled (FZD) in yellow catfish (Pelteobagrus fulvidraco) by copper exposure
投稿时间:2017-04-18  修订日期:2017-07-19
DOI:10.11964/jfc.20170410801
中文关键词: 黄颡鱼  FZD  分子特征  组织表达  
英文关键词: Pelteobagrus fulvidraco  Frizzled (FZD)  molecular characterization  tissue distribution  copper exposure
基金项目:国家自然科学基金(31422056)
作者单位E-mail
张丽晗 华中农业大学水产学院, 动物遗传育种与繁殖教育部重点实验室, 湖北 武汉 430070  
罗智 华中农业大学水产学院, 动物遗传育种与繁殖教育部重点实验室, 湖北 武汉 430070 luozhi99@mail.hzau.edu.cn 
有文静 华中农业大学水产学院, 动物遗传育种与繁殖教育部重点实验室, 湖北 武汉 430070  
李丹丹 华中农业大学水产学院, 动物遗传育种与繁殖教育部重点实验室, 湖北 武汉 430070  
吴坤 华中农业大学水产学院, 动物遗传育种与繁殖教育部重点实验室, 湖北 武汉 430070  
潘亚雄 华中农业大学水产学院, 动物遗传育种与繁殖教育部重点实验室, 湖北 武汉 430070  
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中文摘要:
      实验采用RT-PCR和RACE克隆fzd2、fzd3afzd4和fzd10基因,其cDNA全长分别为2 176、3 243、2 509和3 021 bp,其中ORF长度分别为1 652、2 084、1 649和2 161 bp,编码551、695、550和586个氨基酸。氨基酸序列比对和系统树分析显示,4种基因十分保守,黄颡鱼与斑点叉尾鮰亲缘关系最近。组织表达分析显示,4种基因的mRNA在脑、脾脏、肾脏、鳃、心脏、肌肉、脂肪、肝脏及卵巢等组织中都有表达,而其表达量不尽相同。FZD家族基因对铜的响应研究表明,在暴露28 d时,fzd3a mRNA 水平随着铜浓度的增加而增加,但是fzd2、fzd4和fzd10基因表达各个处理组无显著性差异;在56 d,fzd2 mRNA水平在30 μg Cu/L组最高,其他2个组差异不显著,fzd3afzd4和fzd10的基因表达在3个铜暴露组中均无显著差异,表明fzd家族这些基因的功能发生了分化,部分成员介导了铜影响黄颡鱼卵巢发育的调控。
英文摘要:
      Four complete FZD cDNA sequences, designated fzd2, fzd3a, fzd4 and fzd10, were amplified by RT-PCR and RACE approaches from Pelteobagrus fulvidraco. The validated cDNAs encoding for fzd2, fzd3a, fzd4 and fzd10 were 2 176, 3 243, 2 509 and 3 021 bp in length, encoding peptides of 551, 695, 550 and 586 amino acid residues, respectively. The amino acid alignment and phylogenetic analysis revealed that Fzd sequences were relatively conserved among different vertebrates, and fzds of P. fulvidraco were more closely related to those of Ictalurus punctatus. All fzds mRNAs were expressed in all tested tissues (brain, spleen, kidney, gill, heart, muscle, mesenteric fat, liver and ovary), but at variable levels. On day 28, fzd3a mRNA levels increased with increasing waterborne Cu concentration, but mRNA expression showed no significant differences among the three groups. On day 56, fzd2 mRNA levels were the highest in 30 μg Cu/L group and showed no significant differences between other two groups, and mRNA levels of fzd3a, fzd4 and fzd10 showed no significant differences among three groups. The present study will be helpful to understanding the physiological functions of fzd genes.
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