金钱鱼agrp基因cDNA克隆及饥饿与复投喂对其表达的影响  

Molecular Cloning of the agrp Genes in Scatophagus argus and Their Expression Pattern under the Treatment of Starvation and Refeeding

作　　者：蔡博生 陈华慧 罗翠婷 翟毅 李广丽[1,2] 邓思平 CAI Bosheng;CHEN Huahui;LUO Cuiting;ZHAI Yi;LI Guangli;DENG Siping(Fisheries College,Guangdong Ocean University,Zhanjiang 524088,China;Guangdong Research Center on Reproductive Control and Breeding Technology of Indigenous Valuable Fish Species,Zhanjiang 524088,China)

机构地区：[1]广东海洋大学水产学院,广东湛江524088 [2]广东省名特优鱼类生殖调控与繁育工程技术研究中心,广东湛江524088

出　　处：《水产科学》2023年第5期821-829,共9页Fisheries Science

基　　金：国家自然科学基金资助项目(31972775);大学生创新创业计划项目(CXXL2018132)。

摘　　要：为探究刺鼠相关蛋白/神经肽Y (AgRP)神经元在金钱鱼应答饥饿与复投喂过程中所起的调控作用,采用反转录PCR自金钱鱼下丘脑克隆了agrp1和agrp2 2种亚型基因,并采用实时荧光定量PCR分析了2种基因在金钱鱼不同组织中的表达情况和正常投喂组(2、7 d)、饥饿组(2、7 d)、复投喂组(饥饿7 d然后复投喂3 h)金钱鱼(体质量52~60 g)下丘脑、肝脏和肠道中agrp1和agrp2基因的表达情况。序列分析结果显示,agrp1和agrp2基因的开放阅读框长度分别为429 bp和351 bp。组织表达结果显示,agrp1基因主要在下丘脑和肌肉中表达,agrp2基因主要在下丘脑和脑垂体中表达。饥饿及复投喂结果表明:下丘脑中,agrp1基因表达水平在饥饿2 d后差异不显著,饥饿7 d后显著下调,复投喂后其表达水平恢复正常;肝脏中,agrp1基因表达水平在饥饿2 d后下调,但饥饿7 d后显著上调,复投喂后其表达水平下调到正常水平;肠道中,agrp1基因表达水平在饥饿2 d后无显著差异,饥饿7 d后显著上调,复投喂后其表达水平下调到正常水平。饥饿和复投喂期间,下丘脑、肝脏和肠道中agrp2基因表达水平与对照组差异不显著。结果表明,下丘脑、肝脏和肠道中的agrp1基因参与了金钱鱼摄食过程的调节。To investigate the roles of agouti-related protein/neuropeptide Y (AgRP) neurons in response to starvation and refeeding in spotted scat Scatophagus argus . Agrp1 and agrp2 genes were cloned from the hypothalamus of spotted scat by RT-PCR. The expression level of the two genes in different tissues was detected by qPCR. The expression of agrp1 and agrp2 genes in the hypothalamus, liver and intestine of the normal feeding group (2 d, 7 d), starvation group (2 d, 7 d), and refeeding group (7 d starvation followed by refeeding for 3 h) of spotted scat (body weight: 52—60 g)was investigated, respectively. Sequence analysis showed that the open reading frames of agrp1 and agrp2 genes were 429 bp and 351 bp, respectively. Agrp1 gene was mainly expressed in hypothalamus and muscle, whereas agrp2 gene was mainly expressed in hypothalamus and pituitary. It showed that there was no significant change was observed in the expression level of agrp1 gene in hypothalamus after 2 d of food deprivation. However, it decreased after 7 d of food deprivation and then increased to the normal level after refeeding. In liver, the expression level of agrp1 gene decreased after 2 d of food deprivation, but increased after 7 d of food deprivation and then decreased to the normal level after refeeding. In intestine, the expression level of agrp1 gene did not change after 2 d of food deprivation. However, it increased significantly after 7 d of food deprivation and then decreased to the same levels of the control group after refeeding. In terms of the expression level of arrp2 gene in the hypothalamus, liver and intestines of the starved group, there was no significant difference comparing with the control group during the fast feeding and refeeding period. In summary, the agrp1 gene might be involved in the regulation of food intake in hypothalamus, liver and intestine in spotted scat.

关 键 词：金钱鱼 刺鼠相关蛋白 饥饿及复投喂 摄食 

分 类 号：Q786[生物学—分子生物学]