J Korean Soc Pediatr Nephrol.  2011 Oct;15(2):138-145.

The Change of Podocyte beta-Catenin by Puromycin Aminonucleoside

• Choi JY
• Ahn EM
• Park HY
• Shin JI
• Ha TS

Abstract

PURPOSE
To test whether the expression of beta-catenin, a component of podocyte as a filtration molecule, would be altered by puromycin aminonucleoside (PAN) in the cultured podocyte in vitro.
METHODS
We cultured rat glomerular epithelial cells (GEpC) with various concentrations of PAN and examined the distribution of beta-catenin by confocal microscope and measured the change of beta-catenin expression by Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR).
RESULTS
We found that beta-catenin relocalized from peripheral cytoplasm to inner cytoplasm, therefore, intercellular separations were seen in confluently cultured cells by high concentrations of PAN in immunofluorescence views. In Western blotting of GEpC, PAN (50 microg/mL) decreased beta-catenin expression by 34.9% at 24 hrs and 34.3% at 48 hrs, compared to those in without PAN condition (P<0.05). In RT-PCR, high concentrations (50 microg/mL) of PAN also decreased beta-catenin mRNA expression similar to protein suppression by 25.4% at 24 hrs and 51.8% at 48 hrs (P<0.05).
CONCLUSION
Exposure of podocytes to PAN in vitro relocates beta-catenin internally and reduces beta-catenin mRNA and protein expression, which could explain the development of proteinuria in experimental PAN-induced nephropathy.

Keyword

beta-Catenin; Puromycin aminonucleoside (PAN)-induced nephropathy; Glomerular epithelial cells (GEpC); Podocyte

MeSH Terms

Animals
Ascorbic Acid
beta Catenin
Blotting, Western
Cells, Cultured
Cytoplasm
Epithelial Cells
Filtration
Fluorescent Antibody Technique
Glycyrrhetinic Acid
Podocytes
Proteinuria
Puromycin
Puromycin Aminonucleoside
Rats
RNA, Messenger
Ascorbic Acid
Glycyrrhetinic Acid
Puromycin
Puromycin Aminonucleoside
RNA, Messenger
beta Catenin