刀鲚POMC基因的cDNA克隆及其应激应答  

Cloning of POMC cDNA and the stress response in Coilia nasus

作　　者：杜富宽[1,2] 徐钢春[2] 黎燕[2] 练青平[1] 徐跑[2] 

机构地区：[1]农业部淡水渔业健康养殖重点实验室,浙江省淡水水产遗传育种重点实验室,浙江省淡水水产研究所,浙江湖州313001 [2]中国水产科学研究院淡水渔业研究中心,农业部淡水渔业和种质资源利用重点实验室,江苏无锡214081

出　　处：《中国水产科学》2017年第2期231-238,共8页Journal of Fishery Sciences of China

基　　金：农业部淡水渔业健康养殖重点实验室开放课题(ZJK2014003);中国水产科学研究院淡水渔业研究中心中央级公益性科研院所基本科研业务费专项资金(2015JBFR06);江苏省科技支撑项目(BE2014307)

摘　　要：为了研究刀鲚(Coilia nasus)阿黑皮素原(proopiomelanocortin,POMC)基因的应激应答及调控,克隆了刀鲚POMC基因的c DNA全长。刀鲚POMC基因的c DNA全长2030 bp,编码区699 bp,编码232个氨基酸。二级结构预测显示,刀鲚POMC包括信号肽(signal peptide)、N末端区(N-terminal region,NPP)、促肾上腺皮质激素结构域(adrenocorticotropic hormone,ACTH)、β-脂解素(β-LPH)、α-促黑素(α-MSH)、促肾上腺皮质激素样垂体中叶多肽(CLIP)、γ-脂解素(γ-LPH)、β-促黑素(β-MSH)和β-内咖肽(β-EP)结构域。运用荧光定量的方法检测了该基因在刀鲚不同组织中的表达分布。结果显示,POMC在健康刀鲚的脑高表达,鳃、肾、精巢相对高表达,肝、脾、肠、头肾、肌肉和卵巢中微量表达。运输胁迫后,POMC基因的表达量显著下降(P<0.05);10‰ NaCl组4 h和6 h POMC基因的表达量显著上调(P<0.05),6 h的表达量恢复至应激前。POMC基因在脂肪代谢及应激调控中发挥重要的功能,本研究结果可为该基因在刀鲚后续育种及应激调控工作提供重要基础。The estuarine tapertail anchovy（Coilia nasus, junior synonym C. ectenes） is widely distributed in the Yangtze River, the coastal waters of China and Korea, and the Ariake Sound of Japan. It is a commercially important species because of its nutritive value and popularity. However, excess fishing and changes in aquatic ecology have almost caused extinction of this species in the middle reaches of the Yangtze River. In recent years, several studies have been performed on pond breeding and artificial propagation, and larval rearing techniques have been established. Thus, the threat to C. nasus resources has been alleviated. However, C. nasus is very sensitive to stress, which often results in death and huge economic losses. Little is known about the mechanism by which loading stress causes death in fish. Pro-opiomelanocortin（POMC） is synthesized in the corticotrophs and melanotrophs of the anterior and intermediate lobes of the pituitary, respectively, as well as in peptidergic neurons in the arcuate nucleus of the hypothalamus. It is post-translationally cleaved into peptide hormones, including adrenocorticotrophic hormone, β-endorphin, α-, β-, and γ-melanocyte stimulating hormone（MSH）, N-POMC, and β-lipotropin（β-LPH） in tissue- and cell dependent manners. These peptides exhibit different physiological functions, such as promoting ovarian maturation, lipolysis, inhibiting fat accumulation, regulating osmotic pressure, and reacting to stressors. We cloned the POMC gene in C. nasus and determined that the full-length c DNA is 1212 bp long, with a 699 bp open reading frame that encoded a 232 amino acid protein. The POMC protein is predicted to contain the typical POMC domains of a signal peptide sequence, the N-terminal region, and the adrenocorticotrophic hormone, β-LPH, α-MSH, CLIP, γ-LPH, β-MSH, and β-EP domains. Gene expression patterns were detected in different tissues by quantitative reverse transcription-polymerasechain reaction. POMC was expressed strongly in the brain and relativel

关 键 词：刀鲚 POMC基因 应激应答 运输胁迫 

分 类 号：S96[农业科学—水产养殖]