锦鲤早期发育过程中色素代谢相关酶的活性和基因表达变化  

Changes of pigment-related enzyme activity and gene expression at early developmental stage of koi carp

作　　者：殷浩然 罗明坤 王兰梅[2] 董在杰[1,2] 朱文彬[2] 傅建军[2] YIN Haoran;LUO Mingkun;WANG Lanmei;DONG Zaijie;ZHU Wenbin;FU Jianjun(Wuxi Fisheries College,Nanjing Agricultural University,Wuxi 214081,China;Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization,Ministry of Agriculture and Rural Affairs,Freshwater Fisheries Research Centre of Chinese Academy of Fishery Sciences,Wuxi 214081,China)

机构地区：[1]南京农业大学无锡渔业学院,江苏无锡214081 [2]中国水产科学研究院淡水渔业研究中心,农业农村部淡水渔业与种质资源利用重点实验室,江苏无锡214081

出　　处：《南方水产科学》2019年第5期109-117,共9页South China Fisheries Science

基　　金：江苏省第五期“333工程”培养资金(BRA2017083);农业农村部现代农业人才支撑计划项目(2016142);中央级公益性科研院所基本科研业务费专项资金(2019ZY22)

摘　　要：通过显微观察、酶联免疫吸附及荧光定量PCR等方法,探究了红白、红黑锦鲤(Cyprinus carpio var.koi)子一代(F_1)早期体色发育过程及不同发育时期色素代谢相关酶(酪氨酸酶、胱氨酸酶)的活性和基因(agouti、mc1r、mitf、tyr)的表达变化。结果表明,锦鲤出膜后呈淡黄色半透明,黑色素细胞在出膜8 d后开始出现。胱氨酸酶活性随锦鲤发育呈升高趋势,出膜7 d后显著升高(P<0.05);7日龄仔鱼前,红黑组与红白组间无显著差异(P>0.05);出膜7 d后红黑组比同时期红白组显著升高(P<0.05)。酪氨酸酶在锦鲤发育过程中也呈升高趋势,但略有起伏,眼色素积累期及出膜23 d后酪氨酸酶活性均比相邻各期高(P<0.05);组间比较发现发育各期红黑组均比红白组高(P<0.05)。表明酪氨酸酶、胱氨酸酶活性变化与锦鲤色素细胞发育存在密切关联。agouti、mc1r、mitf及tyr 4个基因均在原肠期与神经胚期时表达最高(P<0.01),随后均呈下降趋势,表明这4个基因可能在胚胎发育早期的体色形成过程中发挥着重要作用。We investigated the changes of tyrosinase and cystinase activities,as well as the expressions of tyr,mitf,agouti and mc1r mRNAs at the early developmental stage of Kohaku and Hi-Utsuri koi carp(Cyprinus carpio)by using histological observation,enzyme-linked immunoassay and quantitative reverse transcription-polymerase chain reaction methods.The results show that the hatched carps became light yellow and translucent.The melanocytes occurred on 8 dph(day-post-hatching).The activity of cystinase increased with the koi carp development,and no significant difference was observed between Kohaku and Hi-Utsuri groups before 7 dph(P>0.05),while Hi-Utsuri group showed significantly higher enzyme activity than Kohaku group at the same stages after 7 dph(P<0.05).Tyrosinase activity showed an up-regulation trend with slight fluctuation during the development of koi carp,and peaked at the ocular pigment stage and on 23 dph(P<0.05).The enzyme activity was significantly higher in Hi-Utsuri group than in Kohaku group at each developmental stage(P<0.05),which indicates that tyrosinase and cystinase activities were closely associated with pigment cell development of koi carp.The expressions of agouti,mc1r,mitf and tyr mRNAs peaked in gastrula and blastula periods(P<0.01),then decreased.Thus,agouti,mc1r,mitf and tyr mRNAs might play an important role in the formation of body colors in early embryonic development.

关 键 词：锦鲤 体色 酪氨酸酶 胱氨酸酶 tyr MITF AGOUTI MC1R 

分 类 号：S965.8[农业科学—水产养殖]