【摘要】 目的明确新生小牛视网膜视杆-双极细胞体外发育过程中蛋白激酶Cα亚型(PKCα)的表达规律。方法分离培养新生小牛视网膜神经细胞,培养10、20、30和40 d的神经元用于免疫细胞化学染色,采用PKCα抗体标记视网膜视杆-双极细胞,用图像分析系统定量测量阳性细胞的免疫反应强度。结果培养10 d时,PKCα阳性神经元只有1~2个较短的神经突起;到培养20 d时,阳性细胞的一端为1个细长的神经突起,相对的另一端为丛状的短的树状突起,培养30 d及40 d的神经元保持培养20 d时神经元的形态特征。不同培养阶段的阳性细胞中PKCα免疫反应物质均位于细胞体和神经突起,定量分析显示培养20 d的阳性细胞免疫反应强度最高,培养30 d和40 d时阳性细胞PKCα免疫反应强度下降。结论培养20 d的视杆-双极细胞具备细胞形态和免疫反应特性上的成熟特征,PKCα的表达水平最高,为发育成熟的神经元。培养30 d以后,视杆-双极细胞逐渐衰老。实验结果为选择合适的供体材料进行视网膜移植提供了理论依据。更多还原

【Abstract】 ObjectiveTo investigate the expression of protein kinase Cα in developing neonatal bovine retinal rod-bipolar cells in vitro.MethodsNeonatal bovine retinal cells were dissociated and cultured for 10,20,30 and 40 days in vitro.Retinal rod-bipolar cells were identified by PKCα antibody,and the immunoreactivity of stained cells was measured by image analysis system.ResultsImmunoreactive cells cultured for 10 days possessed 1 or 2 short neurites.Rod-bipolar cells grown for 20 days in vitro extended a long process on one pole of the cell body and a cluster of dendritic processes on the opposite pole.The morphology of stained neurons grown for 30 days and 40 days in vitro was the same as that of 20-day cells.Neurons in different culture periods showed immunostaining both in cell body and neurites.The immunoreactivity(OPTDM) of cells was 0.39±0.049 after cultured for 10 days and had the highest immunoreactivity(0.451±0.054)in 20-day cultured cells.The immunoreactivity of cells cultured for 30-day was 0.206±0.039,and that of 40 days was 0.184±0.028 with a significant decrease in comparison with that of 20 days cultures.ConclusionThe retinal rod-bipolar cells grown for 20 days in vitro have mature morphological and immunoreactive characteristics and the highest expression of PKCα.Rod-bipolar cells after 30 days in vitro tend to undergo degeneration.This study offers a basis for the choice of retinal transplantation donor.更多还原