荧光示踪法在检测神经元细胞间缝隙连接功能中的应用

目的:建立细胞荧光示踪法定量检测神经元细胞间缝隙连接功能。方法:原代培养神经元细胞,免疫荧光法鉴定神经元细胞的纯度。2种不同分子量的荧光染料Calcein和Dil与神经元细胞共同孵育30 min制成供体神经元细胞,与受体神经元细胞共同孵育2.5 h后,在倒置荧光显微镜下观察供体神经元细胞周围的受体神经元细胞数目。结果:原代培养的神经元细胞纯度达90%,可用于检测神经元细胞缝隙连接功能的实验。所建立的方法可以清晰观察到供体神经元细胞周围受体神经元细胞,并可以计数。结论:本实验所建立的荧光示踪法能够定量检测神经元细胞间缝隙连接功能。

Applications of fluorescent tracer method in the detection of neuronal gap junctions

Objective:To establisha method used for quantitative detecting the neuronal gap junction function. Methods:The primary neurons were cultured, the purity of neurons was identified by immunofluorescence. Two kinds of different molecular weight of fluorescent dyes Calcein and Dil were incubated with neurons for 30 min to make donor neurons, and then incubated with received neurons for 2. 5 h. The number of received neurons around the donor neurons was observed with inverted fluorescence microscope. Results:The purity of cultured primary neurons reached 90%, which could be used to defect neurons gap junction function. The received neurons around the donor neurons could be clearly observed and counted. Conclusions:The fluorescent tracer method established in this experiment can be used for quantitative detecting the neuronal gap junction function.