TRPV3在单眼形觉剥夺性弱视大鼠视网膜节细胞中的表达变化

The Change of Expression of TRPV3 in Retinalganglion Cells of Monocular Form-Deprivation Amblyopiarats

目的检测视觉发育敏感期视网膜节细胞中瞬时受体蛋白香草酸亚型3(transient receptor potential vanilloid 3,TRPV3)蛋白的表达变化,探讨其在弱视发病机制中的可能作用。方法2周龄SD大鼠随机分为正常对照组和实验组,实验组缝合大鼠单侧眼睑(左侧)4周,建立剥夺性弱视模型。苏木精-伊红染色(HE染色)观察视网膜各层,尤其是节细胞层的形态学改变。免疫组织化学技术、免疫荧光、蛋白质免疫印迹(western-blot,WB)检测TRPV3蛋白的表达情况。结果实验组缝合侧眼视网膜HE染色与对侧比较:细胞层次结构更紊乱,细胞形态不规则,节细胞层细胞数目变少,细胞核染色更深,这提示单眼形觉剥夺性弱视大鼠视网膜存在细胞损伤或细胞数目的减少。实验组剥夺眼视网膜节细胞与对侧眼视网膜节细胞和正常对照组视网膜节细胞比较,TRPV3蛋白的表达明显减少,差异有统计学意义(P<0.05);实验组未剥夺侧眼视网膜节细胞与正常对照组视网膜节细胞比较,TRPV3蛋白的表达差异无统计学意义(P>0.05)。WB结果与免疫组织化学结果一致。结论视觉发育敏感期内,单眼形觉剥夺后,视网膜节细胞TRPV3蛋白表达降低,可能导致视网膜神经节细胞内Ca2+的运输障碍,从而可能影响了节细胞的生长发育和功能发挥,可能是弱视发生发展的机制之一。

Objective We observed the change of expression of TRPV3 protein in retinal ganglion cells of monocular form-deprivation amblyopia rat within sensitive period of visual development,to explore its possible role in the pathogenesis of amblyopia.Methods Rats aged 2 weeksold were randomly divided into normal control group and experimental group,and unilateral eyelid suture was performed on the experimental group rats for 4 weeks to establish the models of monocular form-deprivationamblyopia.We used IHC,IF,WB analysis to detect the change of expression of TRPV3 protein.Results HE staining:Compared with the contralateral retina of deprived eyes groupthe ipsilateral retinaof deprived eyes group indicates obvious abnormity.The cell layers are disorderly and the cell morphology is irregular and the cell numbers are decreased in the ganglion cell layer.This change indicates that the cell damage or the reduction in the number of cells may happen in the ipsilateral retina of deprived eyes group.IHC and WB results:Compared with the normal control group and the contralateral retinal ganglion cells of deprived eyes group,the expression of TRPV3 protein in ipsilateral retinal ganglion cells of deprived eyes group are remarkable decreased,and the differences are significant(P<0.05).There are no significant differences between normal control group and contralateral retinal ganglion cells of deprived eyes group(P>0.05).Conclusion The expression of TRPV3 proteinis decreasedinthe retinal ganglion cells of monocular form-deprivationamblyopia rats in the sensitive period of visual development,which may cause the transportation abnormity of Ca2+in the ganglion cells of retina,and thereby may affect thedevelopment and function of ganglion cells.This may be one of the path mechanism of occurrence and development of amblyopia.