新生小牛视杆细胞体外发育过程中细胞形态及视色素分子分布的极性特征

目的 明确新生小牛视网膜视杆细胞体外发育过程中细胞形态及视色素分子分布的变化规律。 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rho4D2抗体标记视网膜视杆细胞，观察不同培养时期的阳性细胞的形态特征及视色素分子分布的特点。用图像分析系统定量测量阳性细胞的免疫反应强度。 结果 rho4D2阳性细胞呈现两种形态，一种为无突起的小圆形细胞，另一种为带有顶端突起的细胞。体外培养10 d时，视色素弥漫地分布于整个细胞膜及顶端突起；培养20 d时视色素逐渐向顶端突起或细胞的一端集中；到培养30、40 d时，视色素聚集于顶端突起或细胞的一端，呈现极性分布特征。定量分析显示，体外培养20 d的阳性细胞，免疫反应强度较培养10 d时明显增强，培养20、30、40 d的阳性细胞的免疫反应强度没有明显差异。 结论 体外培养30、40 d的视杆细胞具备细胞结构及视色素分子分布的极性特征，并有高水平的蛋白质表达能力，是发育成熟的神经元。 (中华眼底病杂志, 2005, 21: 394-396)

Cellular configuration and polarized characteristic of visual pigment during the development of rod cells of neonatal calf in vitro

Objective To investigate the changes of cellular configuration and polarized characteristic of visual pigment during the development of rod cells of neonatal calf in vitro. Methods Retinal cells of neonatal calf were dissociated and cultivated for 10, 20, 30 and 40 days in vitro were used for immunocytochemical analysis. Retinal rod cells were identified by rho4D2 antibody. The configuration of positive cells and rhodopsin molecular distribution were analysed at different cultivated time. The immuno-reactivity of positive cells was measured by image analysis system. Results The rho4D2 immuno-reactive cells included small round cells without protuberance and the cells with protuberance at the peak. At the 10th day after cultivation, the visual pigment immunoreaction was suffusive in the whole cell membrane and apical process; while at the 30th and 40th day, it gathered at the membrane of apical process or one pole of the cells. The results of quantitative analysis showed that the immunoreactive intensity of positive cells at the 20th day after cultivation was stronger than that at the 10th day; while there was no significant difference among the immunoreactive intensity at the 20th, 30th, and 40th day. Conclusions Rod cells at the 30th and 40th day after cultivation have the polarized characterization and visual pigment molecular distribution with high level of expressive ability of protein, which are mature neurons.