摘要
目的观察大鼠视网膜神经节细胞(retinal ganglion cells,RGCs)中内源性α晶状体蛋白的表达情况。方法原代培养大鼠RGCs,取胰酶消化后即刻的细胞以及原代培养1～7d的细胞行免疫荧光染色和激光共聚焦显微镜观察,并通过RT-PCR方法检测原代培养1～7d、9d、12d的RGCs中内源性α晶状体蛋白mRNA的表达情况。结果免疫荧光染色结果显示在原代培养后2d的RGCs中,内源性α晶状体蛋白在少许RGCs膜上表达阳性;原代培养3～5d,内源性α晶状体蛋白表达明显增强并达到高峰,呈"戒环状"分布,然后逐渐降低。mRNA检测结果表明,在RGCs原代培养过程中均有内源性α晶状体蛋白的表达,且在培养第3-5天其表达达到高峰,后逐渐降低。结论大鼠RGCs可表达内源性α晶状体蛋白,在离体培养的RGCs中呈一过性高表达,其表达可能与细胞增殖及细胞活力有关。
Objective To investigate the expression of endogenous α-crystallin in retinal ganglion cells（RGCs） in rats in vitro.Methods RGCs of rats were primary cultured.The cells right after digested with trypsogen and the cells primary cultured from 1 day to 7 days were observed by immunofluorescence staining and confocal laser scanning microscopy,the expression of endogenous α-crystallin in the RGCs with primary culture from 1 day to 7 days,9 days,12 days were detected by RT-PCR.Results Immunofluorescence staining showed that,the positive staining of endogenous α-crystallin was observed in little RGCs membrane at 2 days after primary culture,and reached the peak from 3 days to 5 days after primary culture,which in ring shape,then gradually decreased.The results of mRNA detection showed that,the expression of endogenous α-crystallin were observed in the whole procession of primary culture,reached the peak from 3 days to 5 days,then gradually decreased.Conclusion The endogenous α-crystallin can be expressed in the RGCs of rats with temporary increase,which may be relative to the survival and proliferation of RGCs.
出处
《眼科新进展》
CAS
北大核心
2010年第9期815-818,共4页
Recent Advances in Ophthalmology
基金
国家自然科学基金资助(编号:30872833)
第三军医大学青年创新基金资助(编号:2009XQN30)~~
