Kir2.1通道在牛视网膜胶质细胞中的分布和表达-mRNA和蛋白水平的研究  

EXPRESSION AND LOCALIZATION OF THE INWARDLY RECTIFYING K^+ CHANNELS Kir2.1 IN THE BOVINE RETINA

作　　者：潘爱华[1] 罗学港[1] 

机构地区：[1]中南大学湘雅医学院人体解剖学与神经生物学系,长沙410013

出　　处：《神经解剖学杂志》2003年第3期301-306,共6页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金 (No. 3 0 170 3 0 3 )资助项目

摘　　要：本研究目的为在 m RNA和蛋白质水平探讨 Kir2 .1通道在牛视网膜胶质细胞中的分布和表达。从神经视网膜和视网膜色素上皮分离 m RNA和蛋白质 ,以 RT-PCR,Northern blot,Western blot和免疫荧光组织化学法检测 Kir2 .1m RNA和 Kir2 .1蛋白的表达。结果表明 ,RT-PCR扩增的 Kir2 .1c DNA产物在神经视网膜中强表达 ,在视网膜色素上皮中弱表达 ;Northern斑点杂交的 Kir2 .1c DNA仅在神经视网膜表达 ;Western斑点杂交检测 Kir2 .1蛋白单带在神经视网膜中表达 ;免疫荧光组织化学显示 Kir2 .1蛋白在 Muller细胞中分布 ,并且主要分布在 Muller细胞与神经元相接触的细胞膜区域 ,与 Muller细胞的特异性标记蛋白质抗谷氨酸合成酶抗体双重标记显示其分布特性重叠。在视网膜色素上皮未检测到 Kir2 .1蛋白的免疫活性。上述结果表明 ,Kir2 .1分布在视网膜胶质细胞的 Muller细胞 ,这种分布特性可能与完成 Muller细胞功能 -保持细胞外 K+ 的平衡、促进To examine the expression and localization of the inwardly rectifying K + channels Kir2.1 in the glia cells of bovine retina at the mRNA and protein levels, amplified cDNA products by RT-PCR for Kir2.1 were detected in the bovine neural retina and retinal pigment epithelium (RPE). Northern blot analysis showed that the mRNAs for Kir2.1 were only expressed in the neural retina; no specific band was detected in RPE. At the protein level, a single band for Kir2.1 proteins was detected in the neural retina by Western blot analysis. Immunofluorescence revealed that Kir2.1 immunostaining was expressed at many parts of Mller cells, especially in the membrane domains of Mller cells that contact retinal neurons, i.e., along the two stem processes, over the soma, and in the side branches extending into the synaptic layers. No immunostaining was seen in RPE. Double staining showed that Kir2.1 proteins and glutamine synthetase proteins which are a marker of Mller cell colocalized well. These results reveal that Kir2.1 is localized in the Mller cells, and no Kir2.1 is found in RPE. These data suggests that Kir2.1 may be involved in the transport of K + in the bovine neural retina.

关 键 词：胶质细胞 视网膜 牛 Kir2.1通道 分布 表达 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]