培养的胚胎视网膜神经细胞Ca^(2+)分布与Ca^(2+)通道特征  

Distribution of free Ca^(2+) and characterization of Ca^(2+) channel in the cultured early-stage human embryonic retina

作　　者：刘文文[1] 徐萍[1] 郭强苏[2] 刘军[1] 顾青[1] 黄倩[1] 

机构地区：[1]上海市第一人民医院中心实验室,200080 [2]上海第二医科大学组织胚胎学教研室

出　　处：《中华眼底病杂志》2002年第1期42-45,共4页Chinese Journal of Ocular Fundus Diseases

基　　金：国家自然科学基金资助项目 (39770 2 4 2 )

摘　　要：目的　检测分析培养的胚胎早期视网膜神经细胞游离钙 (Ca2 + )分布与钙 (Ca2 + )通道特征。　方法　体外培养 11～ 15周胎儿视网膜神经细胞 ,在含 Ca2 +与不含 Ca2 +的 Hepes缓冲液中与钙荧光指示剂 Fluo3孵育染色 ,同时加入或不加入异博定、佩尔地平或地塞米松 ,共聚焦显微镜观察记录游离 Ca2 +分布 ,以及受不同浓度 K+刺激后 Ca2 +通道开放与 Ca2 +转移情况。　结果　培养的 11～ 15周胎儿视网膜神经元及神经胶质细胞受 K+ 刺激后均出现明显的 Ca2 + 转移与再分布 ,在缺乏细胞外 Ca2 + 的情况下 ,细胞浆内钙库仍可迅速释放 Ca2 +并转移至细胞核内。异博定、佩尔地平及地塞米松能够抑制细胞外 Ca2 +进入视网膜神经细胞内。　结论　胚胎早期视网膜神经元及神经胶质细胞存在 L型 Ca2 + 通道 。Objective To investigate the distribution of free Ca 2+ and characterization of Ca 2+ channel in the cultured 11～15 week human embryonic retina. Methods The cells from 11～15 week embryonic retina were dissociated enzymatically and cultured on coated cover slides. Three weeks later, Fluo3, a Ca 2+ indicator, was added in Ca 2+ containing or no Ca 2+ Hepes buffer and incubated with retinal cells for 30 min, meanwhile with or without verapamil, perdipine, or dexamethasone adding. Ca 2+ staining and Ca 2+ transit before and after 10 μmol/L or 50 μmol/L KCl stimulation was observed and recorded using Zeiss LM510 confocal laser scanning microscopy. Results In response to K + exposure, a rapid rise of free Ca 2+ in cytoplasma and nuclei of neuron and glial cell was observed, but was repressed with the treatment of verapamil, perdipine, or dexamethasone. The obvious Ca 2+ influx from cytoplasma into nuclei was observed even in the absence of external Ca 2+ . Conclusion L type Ca 2+ channel was expressed and functionally matured in the cultured early stage human embryonic retina.

关 键 词：胚胎 细胞培养 视网膜神经元 钙通道 神经胶质细胞 

分 类 号：R770.2[医药卫生—眼科]