激光共聚焦显微镜和免疫组化染色对比观察KCNQ1在小鼠耳蜗血管纹的表达

Expression and Subcellular Localization of KCNQ1 in Stria Vascularis of Mice Cochlea by Laser Scanning Confocal Microscopy and Immunohistochemistry Staining

目的:采用激光共聚焦显微镜(LSCM)技术和免疫组化染色检测KCNQ1在小鼠耳蜗血管纹中的表达分布特征。方法:以KCNQ1-/-突变纯合子基因型小鼠和CBA/CaJ小鼠为实验对象,采用免疫组化染色、LSCM结合免疫学标记方法对比观察血管纹KCNQ1蛋白表达情况。结果:免疫组化染色显示CBA/CaJ小鼠血管纹边缘细胞顶膜KCNQ1蛋白高表达,呈棕褐色;LSCM观察显示免疫单标KCNQ1绿色荧光蛋白分布于CBA/CaJ小鼠血管纹边缘细胞细胞核,KCNQ1-/-小鼠边缘细胞均未见阳性反应。结论:LSCM技术能准确显示KCNQ1在小鼠耳蜗血管纹中表达及细胞定位。

ObjectiveTo investigate the expression and distribution of KCNQ1 in stria vascularis of murine cochlea with laser scanning confocal microscopy （LSCM） and immunohistochemistry staining technique. Methods： With LSCM and immunohistochemistry staining, the location of KCNQ1 in cochlea of the CBA/CaJ and KCNQ1 -/ -mice were observed. Results： Immunohistochemical staining showed that the expression of KCNQ1 protein was found at the apical membrane wall of marginal cells in stria vascularis of CBA/CaJ mice. There was no KCNQ1 protein expression in marginal cells of KCNQ1 -/-mice. The fluorescence of KCNQl protein was mainly expressed in the cell membrane of marginal cells of CBA/CaJ mice with LSCM. Conclusion：The expression and distribution of KCNQ1 protein in marginal cells of stria vascularis could be observed with LSCM.