Korean J Pediatr.  2012 Oct;55(10):371-376. 10.3345/kjp.2012.55.10.371.

Puromycin aminonucleoside modulates p130Cas of podocytes

• Ha TS
• Choi JY
• Park HY

• KMID: 2140712
• DOI: http://doi.org/10.3345/kjp.2012.55.10.371

Abstract

PURPOSE
Puromycin aminonucleoside (PAN) specifically injures podocytes, leading to foot process effacement, actin cytoskeleton disorganization, and abnormal distribution of slit diaphragm proteins. p130Cas is a docking protein connecting F-actin fibers to the glomerular basement membrane (GBM) and adapter proteins in glomerular epithelial cells (GEpCs; podocytes). We investigated the changes in the p130Cas expression level in the PAN-induced pathological changes of podocytes in vitro.
METHODS
We observed changes in the p130Cas expression in cultured rat GEpCs and mouse podocytes treated with various concentrations of PAN and antioxidants, including probucol, epigallocatechin gallate (EGCG), and vitamin C. The changes in the p130Cas expression level were analyzed using confocal immunofluorescence imaging, Western blotting, and polymerase chain reaction.
RESULTS
In the immunofluorescence study, p130Cas showed a diffuse cytoplasmic distribution with accumulation at distinct sites visible as short stripes and colocalized with P-cadherin. The fluorescences of the p130Cas protein were internalized and became granular by PAN administration in a dose-dependent manner, which had been restored by antioxidants, EGCG and vitamin C. PAN also decreased the protein and mRNA expression levels of p130Cas at high doses and in a longer exposed duration, which had been also reversed by antioxidants.
CONCLUSION
These findings suggest that PAN modulates the quantitative and distributional changes of podocyte p130Cas through oxidative stress resulting in podocyte dysfunction.

Keyword

Cytoskeleton; p130Cas; Podocytes cell; Puromycin aminonucleoside

MeSH Terms

Actin Cytoskeleton
Actins
Animals
Antioxidants
Ascorbic Acid
Blotting, Western
Cadherins
Catechin
Crk-Associated Substrate Protein
Cytoplasm
Cytoskeleton
Diaphragm
Epithelial Cells
Fluorescent Antibody Technique
Foot
Glomerular Basement Membrane
Mice
Oxidative Stress
Podocytes
Probucol
Proteins
Puromycin
Puromycin Aminonucleoside
Rats
RNA, Messenger
Actins
Antioxidants
Ascorbic Acid
Cadherins
Catechin
Crk-Associated Substrate Protein
Probucol
Proteins
Puromycin
Puromycin Aminonucleoside
RNA, Messenger