Molecular detection of heparan sulfate proteoglycan mRNA in rat kidney during calcium oxalate nephrolithiasis

The present study used reverse transcription (RT)-PCR to examine heparan su lfate proteoglycan (HSPG) mRNA expression in rat kidneys. Total mRNA in kid ney was isolated and converted to cDNA. To confirm the exact expression lev el of HSPG mRNA, quantitative competitive (QC)-PCR was performed for each s ample. PCR products were resolved by electrophoresis on 1.5% agarose gel an d visualized with ethidium bromide. Fragment intensity and area were measur ed using an image analyzer. In QC-PCR, target DNA and competitive DNA were expressed, using gene-specific primer for HSPG mRNA, as 506- and 345-bp ban ds, respectively. The level of HSPG mRNA expression apparently increased in the nephrolithic rat kidney. Immunohistochemical study revealed that incre ased production of heparan sulfate was detected in both distal and proximal tubules during nephrolithiasis. These findings suggest that increased expr ession of HSPG may play a significant role during calcium oxalate stone for mation.