PMID: 15388893Sep 25, 2004Paper

Effect of the knockdown of podocin mRNA on nephrin and alpha-actinin in mouse podocyte

Experimental Biology and Medicine
Qingfeng FanJiang-hong Deng

Abstract

Recently, the novel podocyte proteins podocin, nephrin, and alpha-actinin-4 have been identified in three congenital/family nephrotic syndromes, respectively. Further studies showed that these podocyte proteins were involved in some acquired nephrotic syndromes and various experimental models of proteinuria. However, the molecular interactions among these podocyte proteins remain unclear. In this study, to investigate the molecular interactions among podocin, nephrin, and alpha-actinin-4, we reconstructed the RNA interference (RNAi) expression vector, pSilencer 2.1-U6, specifically targeting podocin mRNA, and it was transfected into the mouse podocyte clone (MPC5). Immunofluorescence staining, double-immunolabeling, confocal microscopy, semiquantitative reverse transcription polymerase chain reaction (RT-PCR), and Western blotting were used to detect the distribution and expression of podocin, nephrin, alpha-actinin-4, and glyseraldehyde-3-phosphate dehydrogenase (GAPDH)/beta-actin. The fluorescence intensity of podocin and nephrin decreased obviously, along with the evident distribution change from the cell membrane surface to the nucleus circumference in podocyte. In relation to GAPDH, the mRNA reductions of podocin and nephr...Continue Reading

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Citations

Mar 12, 2013·Evidence-based Complementary and Alternative Medicine : ECAM·Thing-Fong TzengI-Min Liu
Apr 26, 2006·Experimental Biology and Medicine·Yan XingNa Guan
Jun 3, 2011·American Journal of Physiology. Renal Physiology·Seetharamaiah ChittiprolRichard F Ransom
Oct 19, 2012·Virulence·Alfredo Castañeda-RamírezAntonio Verdugo-Rodríguez
Apr 17, 2007·European Journal of Pharmacology·Ghanshyam UpadhyayMahendra Pratap Singh
Jan 25, 2008·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Lan HeYuhao Li
Sep 16, 2008·European Journal of Pharmacology·Ghanshyam UpadhyayMahendra Pratap Singh

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