朱鑫　易潭　陈琳　等

摘要为了解翘嘴鳜胚胎发育中miR146a的表达规律以及温度对miR146a表达的影响，将翘嘴鳜受精卵分别在22 ℃和25 ℃水温下孵化，并利用实时荧光定量方法检测miR146a在各个发育阶段的表达.结果显示，在22 ℃孵化时，miR146a的表达在原肠期前处于较高水平且各时期表达没有显著性差异（P>0.05），在尾芽期后出现显著性降低（P<0.05），并在出膜时达到最低值；在25 ℃孵化时，miR146a的表达直到神经胚期仍处于较高水平且各时期表达没有显著性差异（P>0.05），发育进入尾芽期后表达显著降低（P<0.05），在肌效期达到最低值；miR146a在25 ℃水温孵化时的表达较22 ℃孵化同时期的表达都出现显著降低（P<0.05）.研究表明，miR146a可能主要在胚胎发育后期起到调控胚胎发育的作用，在尾芽期后通过下调表达来促进胚胎的发育；并且温度能影响miR146a的表达，温度升高会降低miR146a在胚胎发育各个时期的表达，将有利于胚胎的发育.

关键词翘嘴鳜；miR146a；发育性表达；胚胎发育；温度调节

中图分类号Q752； S917.4文献标识码A文章编号10002537（2015）05002106

Expression Analysis of Siniperca Chuatsi MiR146a During Embryonic

Development and Effect of Temperature on MiR146a Expression

ZHU Xin 1，2#， YI Tan 1，2#， CHEN Lin1， WU Ping1， WANG Jianhua1，

CHEN Tao2， ZHANG Jianshe 1， CHU Wuying 1*

（1.Department of Biological and Environmental Engineering， Changsha University， Changsha 410003， China；

2. College of Veterinary Medicine， Hunan Agriculture University， Changsha 410128， China）

AbstractTo investigate the expression pattern of miR146a in embryonic developmental stage and the effect of temperature on the expression of miR146a in Siniperca chuatsi， the embryos were incubated under different temperatures and the expression of miR146a during embryonic developmental stage was detected by RTqPCR. The results showed that the expressions of miR146a were at a higher level and there is no significant difference before gastrula stage when the embryos incubated under 22 ℃ （P>0.05）. And then the expressions of miR146a decreased after tailbud stage and decreased to the lowest level at larval stage （P<0.05）. When the embryos incubated under 25 ℃， the expressions of miR146a were at a higher level and there is no significant difference till neurula stage （P>0.05）. Once at tailbud stage， the expression of miR146a significantly decreased （P<0.05） and to the lowest level at muscular effect stage. It was interesting that the expressions of miR146a were significantly decreased at each developmental stage incubated under 25 ℃ when compared with 22 ℃ incubation （P<0.05）. The results indicated that miR146a mainly involved in regulating embryonic development during later embryonic developmental stages and promoted embryonic development by down regulation its expression. In addition， the expressions of miR146a could decrease when the incubation temperature increased， and miR146a might involve in the effect of temperature on embryonic development of Siniperca chuatsi.

Key wordsSiniperca chuatsi； miR146a； developmental expression； embryonic development； temperature regulation

上世纪80年代，随着鳜鱼人工繁殖获得成功，我国开始大规模推广鳜鱼养殖，使之成为我国主要名贵养殖鱼类之一.翘嘴鳜因其肉质细嫩而鲜美，无小刺，且富含蛋白质，现已成为水产品市场的紧俏商品和水产养殖业的重要品种[1].深入探索翘嘴鳜的发育机制能够为鱼类发育生物学研究和水产养殖提供一些有价值的参考资料.

microRNA（miRNA） 是真核生物中一类进化上保守的参与调控基因表达的非编码小分子RNA.它通过与靶基因3′端非翻译区互补配对来促进靶mRNA 降解或翻译抑制，在基因表达中起着负调控作用[23].miRNA在胚胎发育、细胞增殖与分化、肿瘤发生和新成代谢等一系列生物过程中发挥重要的作用[45]，但是有关miRNA在胚胎的表达特性及其在鱼类早期发育中的功能研究还很少.早期的研究表明miRNA对胚胎发育起到重要作用.鼠和人的胚胎干细胞在发育形成胚胎小体时，胚胎干细胞中一些特定的miRNA的表达会下调[67]，缺失miRNA的鼠和斑马鱼胚胎……