Rho-family small GTPases are involved in forskolin-induced cell-cell contact formation of renal glomerular podocytes in vitro

被引:17
作者
Gao, Shuang-yan
Li, Chun-yu
Shimokawa, Tetsuya
Terashita, Takehiro
Matsuda, Seiji
Yaoita, Eishin
Kobayashi, Naoto
机构
[1] Niigata Univ, Grad Sch Med & Dent Sci, Inst Nephrol, Dept Struct Pathol, Niigata, Japan
[2] China Med Univ, Affiliated Hosp 1, Dept Infect Dis, Shenyang 110001, Liaoning, Peoples R China
[3] Ehime Univ, Sch Med, Dept Anat & Embryol, Toon, Ehime 7910295, Japan
关键词
renal glomerulus; podocyte; cell-cell contact; cAMP; Rho-family small GTPases; mouse;
D O I
10.1007/s00441-006-0365-3
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Intercellular adhesions between renal glomerular epithelial cells (also called podocytes) are necessary for the proper function of the glomerular filtration barrier. Although our knowledge of the molecular composition of podocyte cell-cell contact sites has greatly progressed, the underlying molecular mechanism regulating the formation of these cell-cell contacts remains largely unknown. We have used forskolin, an activator of adenylyl cyclase that elevates the level of intracellular cAMP, to investigate the effect of cAMP and three Rho-family small GTPases (RhoA, Cdc42, and Rac1) on the regulation of cell-cell contact formation in a murine podocyte cell line. Transmission electron microscopy and the immunostaining of cell adhesion molecules and actin-associated proteins have revealed a structural change at the site of cell-cell contact following forskolin treatment. The activity of the Rho-family small GTPases before and after forskolin treatment has been evaluated with a glutathione-S-transferase pull-down assay. Forskolin reinforces the integrity of cell-cell contacts, resulting in the closure of an intercellular adhesion zipper, accompanied by a redistribution of cell adhesion molecules and actin-associated proteins in a continuous linear pattern at cell-cell contacts. The Rho-family small GTPases Rac1 and Cdc42 are activated during closure of the adhesion zipper, whereas RhoA is suppressed. Thus, cAMP promotes the assembly of cell-cell contacts between podocytes via a mechanism that probably involves Rho-family small GTPases.
引用
收藏
页码:391 / 400
页数:10
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