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Updated: Oct 18, 2026
Quantitative Real-Time PCR Evaluation of microRNA Expressions in Mouse Kidney with Unilateral Ureteral Obstruction
Published on: August 27, 2020
Molecular detection of heparan sulfate proteoglycan mRNA in rat kidney during calcium oxalate nephrolithiasis
1Department of Urology, Kurume University School of Medicine, Japan. iidas@med.kurume-u.ac.jp
Abstract:
The present study used reverse transcription (RT)-PCR to examine heparan sulfate proteoglycan (HSPG) mRNA expression in rat kidneys. Total mRNA in kidney was isolated and converted to cDNA. To confirm the exact expression level of HSPG mRNA, quantitative competitive (QC)-PCR was performed for each sample. PCR products were resolved by electrophoresis on 1.5% agarose gel and visualized with ethidium bromide. Fragment intensity and area were measured 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 bands, respectively. The level of HSPG mRNA expression apparently increased in the nephrolithic rat kidney. Immunohistochemical study revealed that increased production of heparan sulfate was detected in both distal and proximal tubules during nephrolithiasis. These findings suggest that increased expression of HSPG may play a significant role during calcium oxalate stone formation.
Insights
This study found that heparan sulfate proteoglycan (HSPG) mRNA expression increased in rat kidneys with nephrolithiasis. This suggests HSPG plays a role in calcium oxalate stone formation.
Area of Science:
- Nephrology
- Molecular Biology
- Biochemistry
Background:
- Heparan sulfate proteoglycans (HSPGs) are crucial components of the kidney extracellular matrix.
- Altered HSPG expression is implicated in various kidney diseases, including stone formation.
Purpose of the Study:
- To investigate the expression levels of heparan sulfate proteoglycan (HSPG) mRNA in rat kidneys during nephrolithiasis.
- To determine the localization and production of heparan sulfate in the kidney during stone formation.
Main Methods:
- Reverse transcription (RT)-PCR and quantitative competitive (QC)-PCR were employed to measure HSPG mRNA expression.
- Immunohistochemistry was used to assess heparan sulfate production in kidney tissues.
- Electrophoresis and image analysis were utilized for PCR product quantification.
Main Results:
- HSPG mRNA expression was significantly elevated in nephrolithic rat kidneys compared to controls.
- Immunohistochemical analysis revealed increased heparan sulfate production in both proximal and distal tubules of affected kidneys.
- Specific PCR product bands of 506-bp (target) and 345-bp (competitive) were observed for HSPG mRNA.
Conclusions:
- Increased expression of HSPG mRNA and protein is associated with calcium oxalate nephrolithiasis in rats.
- These findings suggest a potential role for HSPG in the pathogenesis of kidney stone formation.
- Further research into HSPG's function could lead to novel therapeutic strategies for nephrolithiasis.
