间接免疫荧光法观察内向整流性钾通道Kir2.1在大鼠睫状体的分布  

Localization of inwardly rectifier potassium channel Kir2.1 in rat ciliary body with indirect immunofluorescence

作　　者：蒙艳斌[1] 潘爱华[2] 周聪发[2] 

机构地区：[1]湖南湘南学院解剖教研室,湖南省郴州市423000 [2]中南大学湘雅医学院人体解剖学与神经生物学系,湖南省长沙市410013

出　　处：《中国组织工程研究与临床康复》2008年第2期245-247,共3页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：湖南省教育厅资助项目(05C626)~~

摘　　要：目的:Kir2.1通道存在角膜的内皮细胞,参与膜静息膜电位的产生和跨膜转运,睫状体对房水的生成过程中K+的转运起到关键性的作用。文章在蛋白水平探讨Kir2.1在大鼠睫状体的分布情况。方法:实验于2005-03/2006-10在中南大学湘雅医学院人体解剖学与神经生物学系完成。①实验材料:清洁级成年Wistar大鼠8只,雌雄不拘,体质量150～180g,眼睛活动正常,外观正常,角膜透明、无损伤。②实验过程及评估:取正常Wistar大鼠眼球冰冻包埋切片,行苏木精-伊红染色,普通光镜下观察大鼠睫状体的组织结构;在睫状体组织切片加入多克隆兔抗kir2.1一抗,再加AlexaFluor羊抗兔IgG555荧光二抗,用常规间接免疫荧光组织化学方法检测2.1在大鼠睫状体的分布情况。结果:①睫状体组织结构从外向内可分为7层,即睫状体上腔、睫状体肌层、基质层、玻璃膜、色素睫状上皮层、无色素睫状上皮层和内界膜层。②Kir2.1免疫活性分布在睫状体无色素上皮细胞近房水侧的基底面的胞膜和胞浆中,睫状体的色素上皮细胞无免疫活性。结论:Kir2.1分布于大鼠睫状体的无色素上皮的基底面的胞膜和胞浆中,可能与房水的生成有重要关系。AIM： Kir2. 1 existing in corneal epithelial cells can participate in the production of resting membrane potential and transmembrane transport, ciliary body is crucial for the K＋ transportation in hydatoid development. This study is designed to explore the localization of Kit2. 1 in rat ciliary body at protein level. METHODS： The experiment was conducted in the Department of Anatomy and Neurnbiology, Xiangya School of Medicine in Central South University between March 2005 and October 2006.（1）Eight adult Wistar rats of either gender, weighed 150-180 g, with transparent cornea.（2）Normal eyeballs of Wistar rats were embedded frozenly, and hematoxylin-eosin dyeing was applied to observe the histological structure of rat ciliary body on light microscope. After the tissue sections of ciliary body were added with polyclonal rabbit-anti-kit2.1 and Alexa Fluour sheep-anti-rabbit IgG 555, indirect immunofluorescence was used to detect the distribution of Kir2. 1 in rat ciliary body on frozen sections. RESULTS： （1）Histological structure of rat ciliary body included seven layers： supraciliary, ciliary muscle, stroma, lamina vitrea, pigmented ciliary epithelium, unpigmented ciliary epithelium and inner border.（2）Kir2.1 was localized at the basal membrane of non-pigment epithelial cells, but not in pigment epithelial cells. CONCLUSION： The localization of Kir2. 1 in the basal membrane of ciliary unpigmented epithelial cells suggests that it may play an important role in generation of aqueous fluid.

关 键 词：钾通道 睫状体 上皮细胞 组织构建 

分 类 号：R318[医药卫生—生物医学工程]