长时间单色光照射对豚鼠视网膜视锥细胞及其视蛋白表达的影响  

Influence of long-time illumination of monochromatic light on density of cones and opsin expression in guinea pig

作　　者：胡敏[1] 刘永松 刘睿[3] 江春光[1] 褚仁远[3] 袁援生[4] 

机构地区：[1]云南省第二人民医院眼科,昆明650021 [2]昆明市延安医院眼科,650051 [3]复旦大学附属眼耳鼻喉科医院眼科,上海200031 [4]昆明医学院第一附属医院眼科,650032

出　　处：《中华实验眼科杂志》2011年第3期196-201,共6页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金重点项目（30530770）、云南省科技计划项目（2009CA008）

摘　　要：背景面对视觉环境的不断变化，为获得稳定而有效的信息，动物的视觉系统均应根据需要进行必要的调整。色觉系统面临光谱环境的变化，也应该具有一定的可塑性。在长期单色光照射下，视锥细胞密度和视蛋白的表达是否出现相应的变化目前仍不明确。目的研究长时间单色光照射后豚鼠视网膜视锥细胞密度和视蛋白及其mRNA表达的变化。方法30只出生3d的豚鼠按随机数字表法平均分为绿光照射组、紫光照射组和白色7昆合光照射组，分别暴露于530nm的绿光、400nm的紫光及白色混合光照射环境下饲养8周后，制作视网膜铺片，按照对不同波长敏感的光敏视锥细胞在视网膜的分布特点将视网膜分为背侧、腹侧和混合部，分别行免疫细胞化学单标和双标检测，在荧光显微镜下检测不同光敏视锥细胞的密度变化，应用荧光实时定量聚合酶链反应（real-time PCR）法观察不同波长光照射后豚鼠视蛋白mRNA的变化，Western blot法对视蛋白的变化进行定量检测。结果视网膜背侧单标免疫细胞化学检测显示，3组绿敏视锥细胞密度的差异有统计学意义（F=234．28，P〈0．01），与白色混合光照射组相比，绿光照射组绿敏视锥细胞密度明显增多（q=389．68，P〈0．01），紫光照射组明显减少（q=67．11，P〈0．01）。视网膜腹侧单标免疫细胞化学检测显示，3组紫敏视锥细胞的差异无统计学意义（F=3．14，P〉0．05）；视网膜混合区双标免疫细胞化学检测显示，与白色混合光照射组比较，绿光照射组共表达视锥细胞密度明显增多（q=157．55，P〈0．01），紫光照射组共表达视锥细胞密度明显减少（q=254．85，P〈0．01）。与白色混合光照射组相比，绿光照射组绿敏视蛋白及其mRNA的表达均明显增多（q=4．71，P〈0．05；q=184．45，P〈0．01）；紫光照射组长波长敏感视蛋白（L视蛋白�Background The visual system of animal have to optimally adjust in various environmental conditions in order to obtain stable and effective visual function. However, the color vision system of animals which encounter uncertainty of spectral signals should be plastic. Whether the densities of various cones and expression of opsins change with long-time spectral deprivation is unclear. Objective This study was to investigate the changes of cone density as well as the expression of corresponding opsin and mRNA following the long-term illumination of monochromatic light. Methods Thirty 3-day-old guinea pigs were randomized into 3 groups and exposed to the 530 nm green light,400 nm purple light and white light for consecutive 8 weeks respectively. The flat-mounted retinal sample was prepared and divided into dorsal zone, ventral zone and mixed zone anatomically according to the distribution of different light-sensitive cone. The changes in density of cone cells sensitivited to different colored light were detected by single-label or double-label immunocytochemistry. The levels of opsin and its mRNA were determined using Western-blot and real-time PCR respectively. Results The density of green-sensitivity cones was significantly different in the dorsal zone of retina among green light group, purple light group and white light group （ F= 234.28 ,P〈0.01 ）. Compared with white light group,the density of green-sensitive cones in dorsal retina of green light group was obviously higher but that of purple light group was evidently lower （q=389.68,P〈0. 01 ;q=67. 11, P〈0.01 ）. No significant difference was found in the density of purple-sensitive cones in the ventral zone of retina among green light group, purple light group and white light group （ F = 3. 14, P〉0.05 ）. The density of coexpression of the mixed cone cells was increased in green light group （q= 157.55,P〈0. 01 ） but decreased in purple light group （q = 254.85 ,P〈0.01 ） in comparison with white light group. The expression levels of gr

关 键 词：色觉 视锥细胞 视蛋白 绿敏视锥细胞 紫敏视锥细胞 

分 类 号：R77[医药卫生—眼科]