α晶体蛋白与视网膜神经节细胞结合的细胞定位研究

Subcellular localization of interaction between α crystallin and retinal ganglion cells

目的对α-晶体蛋白与视网膜神经节细胞（RGCs）之间的作用进行定位研究。方法原代培养RGCs，Thy1．1免疫荧光法鉴定RGCs并进行纯度检测，将α-晶体蛋白与RGCs共孵育，应用免疫细胞化学方法，对α-晶体蛋白在RGCs上的靶分子进行间接免疫标记，再以光镜、激光共聚焦显微镜和电镜观察标记物，对α-晶体蛋白与RGCs的结合进行亚细胞定位。结果Thy1．1鉴定RGCs纯度为90％～95％。RGCs免疫组化染色结果显示，α-晶体蛋白与其靶分子结合复合物在RGCs膜上呈阳性表达；共聚焦显微镜下发现RGCs有免疫荧光复合物沉积，大量呈“戒指”状分布于细胞膜，少量分布于细胞轴突上。电镜结果显示，RGCs上有免疫复合物沉积，呈“线状”分布在细胞膜上，对照组呈阴性。结论α-晶体蛋白能与RGCs发生结合，其作用靶点在RGCs膜上。α-晶体蛋白与RGCs的结合不是吸附作用，但是否为受体-配体性质的结合还需进一步验证。

Objective To study the subcellular localization of the interaction between α-crystallin on retinal ganglion cells （ RGCs ）. Methods RGCs were cultured primarily and identified by Thyl. 1 antibody, α- crystallin was incubated with RGCs. The target molecules of α-crystallin in RGCs were indirectly labeled by immuocytochemistry and observed with light microscope, laser confocal scanning microscope and electron microscope. Subcellular localization of α-crystallin on RGCs was conducted. Results The purity of RGCs was 90% -95%. The interaction between α-crystallin and RGCs was detected positive. It was observed with confocal laser scanning microscope and electron microscope that there were immune complex deposits on the cell membrane of RGCs, which were distributed like a ring or line. The interaction was negative in Control group. It showed that the interaction occurred between α- crystallin and RGCs. Conclusions α-crystallin can interact with RGCs, with the target towards the membrane of RGCs. The interaction between α crystallin and RGCs is not adsorption, but whether it is a ligand-receptor relation still needs further study.