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Enhanced glomerular profilin gene and protein expression in experimental mesangial proliferative glomerulonephritis
1Department of Biochemistry, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan.
Abstract:
Profilin is a cytoplasmic protein that binds to actin monomer and regulates actin polymerization. In a number of experimental and human glomerular diseases, the mesangial cell expresses alpha-smooth muscle actin and undergoes a phenotypic change to myofibroblast. We used a rat model of mesangial proliferative nephritis induced with antibody to the Thy 1 antigen present on mesangial cells to investigate whether profilin is upregulated. We amplified and sequenced rat profilin cDNA by the reverse-transcribed-polymerase chain reaction (RT-PCR). The nucleotide and amino acid sequences were highly conserved across the mammalian profilins. We raised affinity purified antibody to rat profilin in rabbits immunized with a synthetic profilin peptide (EFTMDLRTKS). At 7 days after disease induction, enhanced expression in both profilin mRNA and protein was demonstrated in the isolated glomeruli by RT-PCR and Western blot analysis. These results suggest that profilin may be involved in the pathogenesis of glomerulonephritis by reorganizing actin cytoskeleton.
Insights
Profilin, a protein regulating actin, is upregulated in rat glomerulonephritis. This suggests profilin
Area of Science:
- Molecular Biology
- Cell Biology
- Nephrology
Background:
- Profilin binds actin monomers, regulating actin polymerization.
- Mesangial cells in glomerular diseases can change to myofibroblasts, expressing alpha-smooth muscle actin.
- The role of profilin in mesangial cell phenotypic changes during glomerulonephritis is unclear.
Purpose of the Study:
- To investigate the expression of profilin in a rat model of mesangial proliferative nephritis.
Main Methods:
- A Thy 1-induced rat nephritis model was used.
- Rat profilin cDNA was amplified and sequenced using reverse-transcribed-polymerase chain reaction (RT-PCR).
- Profilin mRNA and protein levels in isolated glomeruli were assessed via RT-PCR and Western blot analysis.
Main Results:
- Nucleotide and amino acid sequences of rat profilin showed high conservation across mammals.
- Enhanced expression of both profilin mRNA and protein was observed in glomeruli at 7 days post-disease induction.
- Antibody to rat profilin was successfully raised for protein detection.
Conclusions:
- Profilin expression is upregulated during the development of experimental glomerulonephritis.
- These findings suggest profilin's involvement in the pathogenesis of glomerulonephritis.
- Profilin may contribute to disease progression by reorganizing the actin cytoskeleton in mesangial cells.