J Korean Soc Pediatr Nephrol.  2013 Oct;17(2):79-85.

Effect of Puromycin Aminonucleoside on Podocyte P-Cadherin

• Ha TS

Abstract

PURPOSE
To test whether the expression of P-cadherin, a component of slit diaphragms between podocyte foot processes, would be altered by puromycin aminonucleoside (PAN) in a cultured podocyte in vitro.
METHODS
Rat glomerular epithelial cells (GEpC) were cultured with various concentrations of PAN. The distribution of P-cadherin was examined with a confocal microscope. Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR) were used to measure the change in P-cadherin expression.
RESULTS
This study found that P-cadherin was concentrated in the inner and peripheral cytoplasm with high concentrations of PAN under immunofluorescence views. Western blotting of GEpC revealed that PAN induced a decrease of P-cadherin in dose- and time-dependent manners. A high dose (50 microg/mL) of PAN decreased P-cadherin expression by 21.9% at 24 h (P<0.05) and 31.9% at 48 h (P<0.01) compared to those without PAN. In RT-PCR, high concentrations (50 microg/mL) of PAN also decreased P-cadherin mRNA expression, similar to protein suppression, by 23.5% at 48 h (P<0.05).
CONCLUSION
Podocytes exposed to PAN in vitro concentrated P-cadherin internally, and reduced P-cadherin mRNA and protein expression. This could explain the development of proteinuria in experimental PAN-induced nephropathy.

Keyword

P-Cadherin; Puromycin aminonucleoside (PAN)-induced nephropathy; Glomerular epithelial cells (GEpC); Podocyte

MeSH Terms

Animals
Ascorbic Acid
Blotting, Western
Cadherins*
Cytoplasm
Diaphragm
Epithelial Cells
Fluorescent Antibody Technique
Foot
Glycyrrhetinic Acid
Podocytes*
Proteinuria
Puromycin Aminonucleoside*
Puromycin*
Rats
RNA, Messenger
Ascorbic Acid
Cadherins
Glycyrrhetinic Acid
Puromycin
Puromycin Aminonucleoside
RNA, Messenger