Trpc4在原代培养的大鼠肾小球系膜细胞的定位表达

Location of Trpc4 in Primary Cultured Rat Glomerular Mesangial Cells

目的:研究Trpc4原代培养的肾小球系膜细胞中的定位表达。方法:以用筛网法分离大鼠肾小球,培养于添加了20%胎牛血清的DMEM培养液中,倒置显微镜下观察纯化的系膜细胞的形态,电镜下观察细胞的微丝结构;采用免疫荧光检测Trpc4的表达。超速离心法分别提取胞浆和胞膜蛋白,通过Western blot,检测Trpc4在胞膜和胞浆的表达。结果:系膜细胞主要呈梭形,电镜下可见胞浆内微丝结构丰富;胞浆和胞膜均有Trpc4表达,但胞膜表达丰度明显高于胞浆。结论:原代培养的大鼠肾小球系膜细胞表达Trpc4,且主要分布于细胞膜。

Objective：To investigate the location of Trpc4 in primary cultured rat glomerular mesangial cells.Methods：Rat glomeruli were isolated by the different size of meshes and cultured in DMEM with 20% of FBS until glomerular mesangial cells were confluent.To observe the morphology of cells,as well as ultrustructures by transelectronic microscopy such as microfiliaments.Immunofluorecent stainings were performed to investigate location of Trpc4 in cells.Using ultrocentrigufation to extract the proteins from cytosol and plasma membrane respectively followed by western blot analysis for Trpc4 in the two compartments accordingly.Results：Glomerular mesangial cells were mainly characteristic of spindle like cells and microfiliaments structures are aboundant.Trpc4 were detectable in both cytosol and plasma membrane,while the levels were remarkably dominant on the plasma membrane.Conclusion：Trpc4 distributed dominantly on the plasma membrane in primary cultured rat mesangial cells.