Department of Anatomy and Cell Biology, Universitätsmedizin Greifswald, Friedrich-Loeffler-Str. 23c, 17487 Greifswald, Germany.
Am J Physiol Renal Physiol. 2012 May 15;302(10):F1265-77. doi: 10.1152/ajprenal.00468.2011. Epub 2012 Feb 15.
CD151, a member of the tetraspanin family of membrane proteins, is crucially involved in the formation of the glomerular filtration barrier in humans and mice. However, the role of CD151 in podocytes has not been investigated so far. In the present study, we utilized a conditionally immortalized mouse podocyte cell line to characterize CD151 in podocytes and to examine the consequences of manipulating CD151 expression levels. Mouse podocytes endogenously express CD151 as determined by RT-PCR and Western blotting. GFP-CD151 fusion protein localized to the cell membrane, to cell protrusions and cell-cell contacts, colocalizing with actin, β(1)-integrin, zonula occludens-1, and CD9. The expression of GFP-CD151 in cultured podocytes resulted in a marked increase in the presence of thin arborized protrusions (TAPs). TAPs are distinct from filopodia by increased length, protein composition, branched morphology, and slower dynamics. Furthermore, the migration rate of pEGFP-CD151-transfected podocytes was reduced in a wound assay. Fluorescence recovery after photo bleaching measurements revealed a half-time of 3 s for GFP-CD151 consistent with a high mobility of CD151 in the membrane and cytosol. CD151 knockdown in podocytes reduced β(1)-integrin expression and podocyte cell area, indicating diminished adherence and/or spreading. Our results indicate that CD151 importantly modulates podocyte function.
CD151 是四跨膜蛋白家族的成员,在人和小鼠的肾小球滤过屏障的形成中起着至关重要的作用。然而,到目前为止,CD151 在足细胞中的作用还没有被研究过。在本研究中,我们利用条件永生化的小鼠足细胞系来描述 CD151 在足细胞中的作用,并研究操纵 CD151 表达水平的后果。RT-PCR 和 Western blot 检测结果表明,小鼠足细胞内源性表达 CD151。GFP-CD151 融合蛋白定位于细胞膜、细胞突起和细胞-细胞接触处,与肌动蛋白、β(1)-整合素、封闭蛋白-1 和 CD9 共定位。在培养的足细胞中表达 GFP-CD151 会导致薄树突状突起(TAP)的大量出现。TAP 与丝状伪足的区别在于长度增加、蛋白组成、分支形态和较慢的动力学。此外,在划痕实验中,转染 pEGFP-CD151 的足细胞的迁移率降低。光漂白恢复荧光测量显示 GFP-CD151 的半衰期为 3 秒,这表明 CD151 在膜和细胞质中的流动性很高。在足细胞中敲低 CD151 会降低 β(1)-整合素的表达和足细胞的面积,表明粘附和/或铺展能力下降。我们的结果表明,CD151 重要地调节足细胞的功能。