CDK5对羊驼皮肤黑色素细胞TYR和MITF mRNA表达的调节  

Regulation of TYR and MITF mRNA Expression by CDK5 in Alpaca Melanocytes

作　　者：张瑞娜[1] 范瑞文[1] 程志学[1] 田雪[1] 刘佳[1] 高磊[1] 马峥[1] 董常生[1] 

机构地区：[1]山西农业大学动物科技学院,太谷030801

出　　处：《畜牧兽医学报》2011年第12期1712-1717,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(30571070);山西省青年科技研究基金(2008021038-3)

摘　　要：为了证实CDK5是否参与羊驼毛色的形成,本研究主要对CDK5在羊驼黑色素细胞中调节TYR和MITF的表达进行了研究。本研究首先采用免疫组织化学方法检测CDK5在羊驼皮肤黑色素细胞中的定位,再通过脂质体将CDK5转染于羊驼皮肤黑色素细胞,之后通过Western blot和qRT-PCR的方法检测转染后黑色素细胞中CDK5蛋白、TYR和MITF mRNA的表达。免疫组化结果显示CDK5位于黑色素细胞的胞质和细胞核内;West-ern blot结果显示转染组黑色素细胞中CDK5蛋白表达量明显高于对照组;qRT-PCR结果显示CDK5可下调MITF的表达,同时上调TYR的表达,转染组黑色素细胞中MITF和TYR mRNA的表达水平分别是对照组细胞的0.264 9和3.931 3倍。结果揭示CDK5可能通过调节黑色素细胞核中TYR和MITF mRNA的表达,从而参与调控羊驼毛色形成。In order to investigate whether CDK5 could participate in the coat color formation of alpaca,the regulation of CDK5 on TYR and MITF expression in alpaca melanocytes was tested in this study.The localization of CDK5 in cultured alpaca melanocytes was examined by immunohistochemistry.Then CDK5 was transfected into cultured alpaca melanocytes by liposomes.Following the transfection,CDK5 protein was detected by Western blot and the expression of MITF and TYR was detected by quantitative real-time PCR（qRT-PCR）.The immunohistochemistry results showed that CDK5 was expressed in cytoplasm and nucleus of cultured alpaca melanocytes;Western blot results showed that the level of CDK5 protein was apparently higher in transfected melanocytes than that in normal melanocytes,and qRT-PCR results showed that the mRNA abundance of MITF was down-regulated while the mRNA abundance of TYR was up-regulated by the transfection with CDK5,and the expression level of MITF and TYR mRNA in transfected melanocytes was 0.264 9 and 3.931 3 folds higher than that in controlled melanocytes respectively.The results suggest that CDK5 participate in the formation of coat color by regulating TYR and MITF expression in alpaca melanocytes.

关 键 词：CDK5 TYR MITF 黑色素细胞 毛色 羊驼 

分 类 号：S813.3[农业科学—畜牧学]