Blumenthal A, Giebel J, Warsow G, Li L, Ummanni R, Schordan S, Schordan E, Klemm P, Gretz N, Endlich K, Endlich N
Department of Anatomy and Cell Biology, Universitätsmedizin Greifswald, Greifswald, Germany;
Department of Pathology, University Medical Center Göttingen, Göttingen, Germany;
Am J Physiol Renal Physiol. 2015 Mar 15;308(6):F602-13. doi: 10.1152/ajprenal.00190.2014. Epub 2014 Dec 10.
Elevated glomerular pressure represents a high risk for the development of severe kidney diseases and causes an increase in mechanical load to podocytes. In this study, we investigated whether mechanical stress alters gene expression in cultured podocytes using gene arrays. We found that tetraspanin CD9 is significantly upregulated in cultured podocytes after mechanical stress. The differential expression of CD9 was confirmed by RT-PCR and Western blotting under stretched and unstretched conditions. Furthermore, mechanical stress resulted in a relocalization of CD9. To get an insight into the functional role of CD9, podocytes were transfected with pEGFP-CD9. The expression of CD9 induced the formation of substratum-attached thin arborized protrusions. Ca(2+) depletion revealed that podocytes overexpressing CD9 possess altered adhesive properties in contrast to the control transfected cells. Finally, elevated CD9 expression increased migration of podocytes in a wound assay. In summary, our results suggest that upregulation of CD9 may play an important role in podocyte morphology, adhesion, and migration.
肾小球压力升高代表着发生严重肾脏疾病的高风险,并导致足细胞机械负荷增加。在本研究中,我们使用基因芯片研究了机械应力是否会改变培养的足细胞中的基因表达。我们发现,在机械应力作用后,培养的足细胞中四跨膜蛋白CD9显著上调。在拉伸和未拉伸条件下,通过逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹法证实了CD9的差异表达。此外,机械应力导致CD9重新定位。为了深入了解CD9的功能作用,用pEGFP-CD9转染足细胞。CD9的表达诱导了附着于基质的细树枝状突起的形成。钙离子耗竭显示,与对照转染细胞相比,过表达CD9的足细胞具有改变的黏附特性。最后,在伤口实验中,CD9表达升高增加了足细胞的迁移。总之,我们的结果表明,CD9的上调可能在足细胞的形态、黏附和迁移中起重要作用。