PP-1与PP-2A在金鱼眼(球)不同组织/细胞中的表达模式  

Differential Expression Patterns of the Catalytic Subunits for PP-1 and PP-2A in the Ocular Tissues of Goldfish,Carassius auratus

作　　者：刘文彬[1] 胡小慧[1] 惠珊珊[1] 周洁[1] 刘姣[1] 肖亚梅[1] 刘少军[1] 李万程[1,2,3] 

机构地区：[1]湖南师范大学生命科学学院、蛋白质化学与发育生物学教育部重点实验室,长沙410081 [2]Department of Biochemistry and Molecular Biology [3]Department of Ophthalmology and Visual Sciences,University of Nebraska Medical Center,Omaha,Nebraska 68198-5870,USA

出　　处：《中国生物化学与分子生物学报》2010年第12期1135-1142,共8页Chinese Journal of Biochemistry and Molecular Biology

基　　金：美国NIH基金1R01EY015765和1RO1EY01830;教育部长江学者及创新团队计划(No.IRT0445);湖南省芙蓉学者特聘教授基金(No.24030604);国家自然科学基金面上项目(No.30971658);教育厅优秀青年资助项目(No.08B048)资助~~

摘　　要：为确定蛋白磷酸酶-1和-2A(protein phosphatase-1 and phosphatase-2A,PP-1,PP-2A)在水生低等脊椎动物眼球不同组织/细胞中的表达模式,提取金鱼眼球中4种组织视网膜、晶体上皮细胞、晶体纤维和角膜的RNA和蛋白,利用RT-PCR和Western印迹技术进行检测.同时运用荧光免疫组织化学技术进行了定位表达研究.结果表明:1)在mRNA水平上,PP-1催化亚基α和PP-2A催化亚基β在金鱼视网膜、晶体上皮和角膜中表达强烈,与其在高等哺乳类脊椎动物如小鼠中的表达形成鲜明的对比;2)在蛋白水平上,PP-1和PP-2A催化亚基在金鱼视网膜和晶体上皮细胞中的表达较高,而在角膜中的表达最低.这与它们在小鼠眼球视网膜和角膜中同时具备高表达的模式形成对比;3)在金鱼发育48h的眼球中,PP-1和PP-2A催化亚基定位于视网膜的色素视网膜和神经视网膜细胞中,晶体上皮和纤维细胞中及角膜上皮细胞中,而在性成熟金鱼眼球中,此二酶催化亚基主要定位于视网膜的色素视网膜,神经视网膜及晶体上皮细胞中.这些结果为探讨PP-1和PP-2A在金鱼眼球不同组织中的功能打下了重要基础。To explore the differential expression pattern of the catalytic subunits α and β for protein phosphatase-1 and phosphatase-2A（PP-1,PP-2A）in lower vertebrates,we isolated the total RNAs and proteins from four major ocular tissues（retina,lens epithelia,lens fiber and cornea） of goldfish and conducted qualitative analysis using RT-PCR,Western blot and immunohistochemistry.Our results reveal 1） At the mRNA level,PP-1cα and PP-2Acβ display dominant expression in retina,lens epithelia and cornea of goldfish,which is different from the expression pattern in mouse eye where both α and β subunits for PP-1 and PP-2A are strongly expressed;2） At the protein level,PP-1c and PP-2Ac are highly expressed in the retina and lens epithelia but least in the cornea of goldfish,which is contrast to the expression pattern in mouse eye where PP-1c and PP-2Ac are highly expressed in the retina and cornea but least in lens;3） In the 48-hour goldfish embryonic eye,PP-1c and PP-2Ac are localized in both neural retinal and pigment retinal layers,lens epithelial and fiber cells,and cornea;In the adult goldfish,the catalytic subunits for the two phosphatases are mainly found in the neural retinal and pigment retinal layers and lens epithelial cells.These results provide valuable information for our further study of the functions of the two phosphatases in goldfish.

关 键 词：去磷酸化 蛋白磷酸酶-1和-2A(PP-1 PP-2A) 视网膜 晶体 角膜 

分 类 号：Q78[生物学—分子生物学]