IR@PKUHSC  > 北京大学第一临床医学院  > 儿科
学科主题临床医学
Effect of the knockdown of podocin mRNA on nephrin and alpha-actinin in mouse podocyte
Fan, QF; Ding, J; Zhang, JJ; Guan, N; Deng, JH
关键词RNA interference podocyte podocin nephrin alpha-actinin
刊名EXPERIMENTAL BIOLOGY AND MEDICINE
2004-10-01
229期:9页:964-970
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Medicine, Research & Experimental
研究领域[WOS]Research & Experimental Medicine
关键词[WOS]PUROMYCIN AMINONUCLEOSIDE NEPHROSIS ; EXPERIMENTAL MEMBRANOUS NEPHROPATHY ; GLOMERULAR SLIT DIAPHRAGM ; MAMMALIAN-CELLS ; EXPRESSION ; INTERFERENCE ; PROTEIN ; REDISTRIBUTION ; CYTOSKELETON ; COMPONENT
英文摘要

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, serniquantitative 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 nephrin were observed by about 65% and 70%, respectively. The expression of podocin protein was too low to be detected in the interference group. In relation to P-actin, the protein level of nephrin decreased by about 78%. The distribution and the mRNA and protein level of alpha-actinin showed no appreciable change. Alpha-actinin localized mainly in the cytoplasm and also extended to the processes. Thus, the significant decreased expression of nephrin along with the redistribution were detected with the knockdown of podocin mRNA, whereas the expression and distribution of alpha-actinin-4 showed no change. These results suggest that podocin may interact directly with nephrin, but not with alpha-actinin.

语种英语
WOS记录号WOS:000224193900014
Citation statistics
Cited Times:26[WOS]   [WOS Record]     [Related Records in WOS]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/58461
Collection北京大学第一临床医学院_儿科
作者单位Peking Univ, Hosp 1, Dept Pediat, Beijing 100034, Peoples R China
Recommended Citation
GB/T 7714
Fan, QF,Ding, J,Zhang, JJ,et al. Effect of the knockdown of podocin mRNA on nephrin and alpha-actinin in mouse podocyte[J]. EXPERIMENTAL BIOLOGY AND MEDICINE,2004,229(9):964-970.
APA Fan, QF,Ding, J,Zhang, JJ,Guan, N,&Deng, JH.(2004).Effect of the knockdown of podocin mRNA on nephrin and alpha-actinin in mouse podocyte.EXPERIMENTAL BIOLOGY AND MEDICINE,229(9),964-970.
MLA Fan, QF,et al."Effect of the knockdown of podocin mRNA on nephrin and alpha-actinin in mouse podocyte".EXPERIMENTAL BIOLOGY AND MEDICINE 229.9(2004):964-970.
Files in This Item:
There are no files associated with this item.
Related Services
Recommend this item
Bookmark
Usage statistics
Export to Endnote
谷歌学术
谷歌学术Similar articles in
[Fan, QF]'s Articles
[Ding, J]'s Articles
[Zhang, JJ]'s Articles
百度学术
百度学术Similar articles in
[Fan, QF]'s Articles
[Ding, J]'s Articles
[Zhang, JJ]'s Articles
必应学术
必应学术Similar articles in
[Fan, QF]'s Articles
[Ding, J]'s Articles
[Zhang, JJ]'s Articles
Terms of Use
No data!
Social Bookmark/Share
All comments (0)
No comment.
 

Items in the repository are protected by copyright, with all rights reserved, unless otherwise indicated.