摘要
目的探讨形觉剥夺对豚鼠锥细胞视蛋白表达的影响。方法出生1周的豚鼠28只,分为正常对照组和形觉剥夺组,各14只,按自然亮暗周期饲养。形觉剥夺组随机选取一只眼以半透明橡胶遮盖,正常对照组随机选择一眼作为对照。4周后取眼球行RT-PCR检测神经视网膜M-视蛋白和S-视蛋白的mRNA表达,并进行豚鼠视网膜铺片后免疫组织化学方法观察两种视蛋白的分布和表达密度,一抗为兔抗鼠红/绿锥视蛋白抗体或蓝锥视蛋白抗体,二抗为羊抗兔IgG带488荧光抗体。以Leica光学显微镜和Leica共聚焦显微镜观察豚鼠视网膜腹侧(下方)和背侧(上方)及中央区的锥细胞视蛋白的表达。结果实验前屈光状态为:对照眼(5.63±0.87)D,形觉剥夺处理眼(5.47±1.02)D,4周后屈光状态分别为:对照眼(4.38±1.02)D,形觉剥夺处理眼(-3.03±0.78)D;屈光度的改变分别为:(-1.25±0.53)和(-8.38±1.44)D。正常豚鼠的视网膜短波敏感锥细胞分布为腹侧(下方)多于背侧(上方);中央密度最高,密度变化为突变。豚鼠的中波敏感锥细胞分布在背部(上方)多于腹侧(下方),中央密度最高,密度变化为渐变;正常组短波敏感蛋白的表达密度:下方为(805.0±203.3)mm^-2,上方为(100.0±57.7)mm^-2;中央为(1637.2±314.1)mm^-2。形觉剥夺组:下方为(640.9±196.8)mm^-2;中央为:(1016.7±144.6)mm^-2,上方为(70.9±30.8)mm^-2;正常组M-视蛋白的表达密度:上方为(946.2±388.5)mm^-2,中心为(1666.7±137.8)mm^-2,下方为(175.0±100.9)mm^-2;形觉剥夺组:上方为(1436.7±366.0)mm^-2,中心为:(2780.0±180.5)mm^-2,下方为(318.2±172.7)mm^-2。RT-PCR检测示形觉剥夺眼和对照眼的M-视蛋白的相对吸光度值分别为1.06±0.07和0.51±0.10,S-�
Objective To investigate the expression of cone opsins in form-deprived myopia(FDM) of guinea pig. Methods Twenty-eight pigmented guinea pigs aged one-week were randomly divided into two groups: FDM group ( n = 14) and control group ( n = 14). White translucent vinyl diffuser attached to one randomly selected eye was used to deprive eyes of form-vision in FDM group. RT-PCR and immunohistochemistry were performed to compare the opsin mRNA expression and spatial density of photoreceptor opsin in the ventral, central and dorsal retinal areas of two groups using antibody against short and medium-wave-length-sensitive cone opsin. Slides were viewed using Leica confocal microscope and Leica light microscope. Results The distribution of opsin across the guinea pig retinae as determined by antibody labeling of cone opsin was asymmetric, with the dorsal retina dominated by M-opsin, and the ventral retinae by S-opsin. But the ventral area also showed positive staining for the M-opsin. Opsin spatial density decreased with increasing retinal eccentricity for both M and S opsins. The spatial density of S-opsin decreased most steeply for eccentricity up to one-third distance from optic disc to retinal edge. The spatial density of M-opsin gradually decreased with eccentricity. The expression of spatial density of M-opsin was higher than that of S-opsin in all three regions. The expression of density of S-opsin in normal control eye is as follow: (805.0 ±203.3)mm^-2(ventral) ; (100.0 ±57.7) mm^-2 (dorsal) ; (1637. 2 ±314. 1) mm^-2 (Central). FDM eye: (640.9 ±196.8) mm^-2(ventral); (1016.7 ±144.6) mm^-2(central); (70.9 ± 30.8) mm^-2; where The expression of spatial density of M-opsin in normal control eye are: (946.2 ± 388.5)mm^-2(dorsal); (1666.7 ±137.8) mm^-2(central); (175.0±100.9) mm^-2(ventral). FDM eye: ( 1436. 7 ± 366. 0 ) mm^-2 (dorsal) ; ( 2780. 0 ± 180.5 ) mm^-2 (central) ; ( 318. 2 ± 172. 7 ) mm^-2
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2007年第3期266-270,共5页
Chinese Journal of Ophthalmology
基金
国家自然科学基金重点资助项目(30530770)
国家自然科学基金资助项目(30371506)
关键词
近视
形状知觉
视蛋白
豚鼠
Myopia
Form preception
Opsin
Guinea pigs