应用地高辛标记的cRNA探针原位杂交检测胰岛素mRNA

成年雄性Ｗｉｔａｒ大鼠，用地高辛标记的ｃＲＮＡ探针在胰腺切片上进行原位杂交，以检测胰岛Ｂ细胞胰岛素基因表达产物ｍＲＮＡ；同时并用免疫组织化学法显示相邻切片胰岛素免疫反应阳性细胞，进行比较，胰腺经４％多聚甲醋灌注固定，经Ｂｏｕｉｎ液再固定２０ｈ，常石蜡包埋和切片，经胰岛素ｃＲＮＡ探针杂交的胰腺切片中胰岛Ｂ细胞和核仁呈阳性反应，阳性信号为蓝色颗粒，胰岛其他细胞及胰腺泡细胞为阴性，方法照切片中均无阳性信

DETECTION OF INSULIN mRNA BY IN SITU HYBRIDIZATION USING A DIGOXIGENIN-LABELED cRNA PROBE

In situ hybridization using cRNA probe labeled with digoxigenin was performed in sections of pancreas from adult male Wistar rats to detect insulin mRNA in islet B cells.Immunohistochemical method was applied in the adjacent sections to show immunoreactive cells for insulin and in situ comparison with insulin mRNA was performed.The rats were perfused with 4% paraformaldehyde.The pancreata were dissected and postfixed in Bouin's fixative for 20 h.The specimens were embedded in paraffin and sectioned conventionally,The results revealed that in the sections hybridized with insulin cRNA probe,the positive signals were blue in color and located in cytoplasm and in nucleolus of B cells, but the control sections showed negative reaction.The comparison of results for insulin mRNA and insulin immunoreactivity indicated that the positive cells demonstrated by in situ hybridization correspond completely with those by insulin immunohistochemistry.This method is simple and easy to repeat,the results are reliable.