Ueda H, Kobayashi T, Mitsui K, Tsurugi K, Tsukahara S, Ohno S
Department of Anatomy, Yamanashi Medical University, Japan.
Invest Ophthalmol Vis Sci. 1995 Oct;36(11):2318-22.
It has been accepted that retinal dystrophin is localized in the outer plexiform layer (OPL) of the retina, but no electron microscopic evidence has been reported until now. Using immunoelectron microscopy, the authors investigate whether retinal dystrophin localizes at the presynaptic or postsynaptic membrane of synaptic regions.
Monoclonal antibody raised against human dystrophin C-terminus was used. Immunoblotting analysis was used for testing dystrophin protein in retinal tissues. Tissue preparations were stained with the immunofluorescence or immunoperoxidase method and were observed with light and electron microscopy.
Immunoblotting analysis showed a molecular weight of band 420 kDa in the sodium dodecyl sulfate--polyacrylamide gel electrophoresis of the retinal tissues. Immunoelectron microscopy disclosed that retinal dystrophin was localized at the presynaptic membrane of synaptic regions in the OPL.
Rat retinal dystrophin localizes at the presynaptic membrane in the OPL, suggesting that it may play some role in the neurotransmitter release of the photoreceptor cell.
视网膜肌营养不良蛋白定位于视网膜外丛状层(OPL)这一观点已被认可,但迄今为止尚无电子显微镜证据报道。作者运用免疫电子显微镜技术,研究视网膜肌营养不良蛋白是否定位于突触区域的突触前膜或突触后膜。
使用针对人肌营养不良蛋白C末端产生的单克隆抗体。免疫印迹分析用于检测视网膜组织中的肌营养不良蛋白。组织标本采用免疫荧光或免疫过氧化物酶法染色,并通过光学显微镜和电子显微镜观察。
免疫印迹分析显示,在视网膜组织的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中,条带分子量为420 kDa。免疫电子显微镜显示,视网膜肌营养不良蛋白定位于OPL中突触区域的突触前膜。
大鼠视网膜肌营养不良蛋白定位于OPL的突触前膜,提示其可能在光感受器细胞的神经递质释放中发挥某种作用。