细胞色素CYP17a1基因在锦鲤性腺不同发育时期的表达分析  

Expression of Cytochrome CYP17a1 in Different Developmental Phases of Gonad in Koi Carp(Cyprinus carpio)

作　　者：史东杰 谭政 张强 伍仕弘 魏东 李文通 王赛赛 孙砚胜 姜巨峰[6] SHI Dongjie;TAN Zheng;ZHANG Qiang;WU Shihong;WEI Dong;LI Wentong;WANG Saisai;SUN Yansheng;JIANG Jufeng(Fisheries Research Institute,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100068,China;Key Laboratory of Urban Agriculture(North China),Ministry of Agriculture and Rural Affairs,P.R.China,Beijing 100097,China;Fisheries College of Tianjin Agricultural University,Tianjin 300380,China;Beijing Tongzhou District Animal Epidemic Disease Prevention and Control Center,Beijing 101101,China;Beijing Yashi Koi Fish Breeding Technology Limited Company,Beijing 102615,China;Tianjin Fisheries Research Institute,Tianjin Ornamental Fish Technology and Engineering Centre,Tianjin 300221,China)

机构地区：[1]北京市农林科学院水产科学研究所,北京100068 [2]农业农村部华北都市农业重点实验室,北京100097 [3]天津农学院水产学院,天津300380 [4]北京市通州区动物疫病预防控制中心,北京101101 [5]北京雅仕锦鲤养殖技术有限公司,北京102615 [6]天津市水产研究所暨天津市观赏鱼技术工程中心,天津300221

出　　处：《河南农业科学》2022年第2期139-146,共8页Journal of Henan Agricultural Sciences

基　　金：北京市观赏鱼产业技术体系创新团队建设专项(BAIC03);北京市农业农村局、北京市财政局、北京市园林绿化局联合攻关项目(JZNF20210610);北京市农林科学院水产科学研究所科研事业经费项目(BJSCS-BZ-2021);天津市海水养殖产业技术体系海水观赏水族岗位项目(ITTMRS2021004);天津市水产研究所观赏鱼创新团队项目(SCYJS201902)。

摘　　要：为探究CYP17a1基因在锦鲤(Cyprinus carpio)性腺不同发育时期的作用及其在不同组织中的表达水平,采用实时荧光定量PCR和蛋白质免疫印迹(Western blot)技术,分析了细胞色素CYP17a1基因在1~3龄锦鲤成鱼性腺发育6个时期(Ⅰ期、Ⅱ期、Ⅲ期、Ⅳ期、Ⅴ期、Ⅵ期),及精巢(卵巢)、脑、肌肉和肾脏等不同组织中CYP17a1 mRNA和蛋白质的表达情况。结果显示,1~3龄锦鲤雌鱼、雄鱼性腺发育不同时期成熟系数(GSI)变化规律基本相同,其中3龄锦鲤性腺Ⅴ期时最高,但卵巢GSI明显高于精巢。CYP17a1 mRNA在卵巢、精巢、脑、肌肉和肾脏都有表达,各组织表达量存在显著差异(P<0.05),表现为精巢>卵巢>肌肉>肾脏>脑,卵巢、精巢的表达量随着年龄增加有升高趋势,在脑、肌肉和肾脏中无明显年龄差别。CYP17a1基因在1~3龄锦鲤卵巢、精巢的6个不同发育时期表达量均值呈现先降低后升高趋势,并呈现出随年龄增长,表达量显著升高的现象。3龄锦鲤精巢Ⅰ期表达量最高,是同龄锦鲤精巢Ⅲ期和Ⅳ期表达量的2.75倍和3.13倍。Western blot检测结果显示,CYP17a1蛋白表达水平与mRNA表达水平一致。以上研究结果表明,细胞色素CYP17a1基因在锦鲤性腺发育中发挥着一定作用,可能在精巢发育及精子细胞分化成熟方面具有重要促进作用。To explore the role of the CYP17a1 gene in different developmental stages of koi fish(Cyprinus carpio)gonads and its expression levels in different tissues,real-time PCR and Western blot were used to analyze the expression of CYP17a1 mRNA and protein of 1—3 years old adult fish in different tissues at six stages(stageⅠ,stageⅡ,stageⅢ,stageⅣ,stageⅤ,stageⅥ).Our results showed that the various rules of maturity index(GSI)of the gonads of 1—3 years old koi females and males at different stages were basically the same.Among them,the 3-year-old koi had the highest GSI value at stage V of gonad,but the ovarian GSI value was significantly higher than that of the testis.The CYP17a1 mRNA was expressed in the ovary,testis,brain,muscle and kidney,respectively.There were significant differences in the expression levels of each tissue(P<0.05),which showed as testis>ovary>muscle>kidney>brain.The expression levels of the ovary and testis increased with age,but there was no significant age difference in brain,muscle,and kidney.The mean expression of the CYP17a1 gene in ovary and testis of 1—3 years old koi at six different developmental stages showed a trend of first decreasing and then increasing,and the expression increased significantly with age.The expression level of testisⅠof 3-year-old koi was the highest,which was 2.75 times and 3.13 times the expression of stageⅢandⅣof the same age koi,respectively.The Western blot test results showed that the expression level of the CYP17a1 protein was consistent with the expression level of mRNA.Our results indicate that the cytochrome CYP17a1 gene plays an important role in promoting the development of koi testis and sperm cell differentiation and maturation.

关 键 词：锦鲤 CYP17a1 卵巢 精巢 表达 实时荧光定量PCR 

分 类 号：S965.1[农业科学—水产养殖]