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Updated: Jul 20, 2026

Single-channel Analysis and Calcium Imaging in the Podocytes of the Freshly Isolated Glomeruli
Published on: June 27, 2015
Synaptic vesicle protein 2B is expressed in podocyte, and its expression is altered in proteinuric glomeruli
Naoko Miyauchi1, Akira Saito, Tamaki Karasawa
1Department of Cell Biology, Institute of Nephrology, Niigata University Graduate School of Medical and Dental Sciences, 1-757 Asahimachi-dori, Niigata 951-8510, Japan.
Abstract:
Synaptic vesicle protein 2B (SV2B) was identified by the subtraction hybridization technique as a molecule of which mRNA expression was decreased in puromycin aminonucleoside (PAN) nephropathy by glomerular cDNA subtraction assay. The expression of SV2B was detected in glomerular lysate with Western blot analysis. Dual-labeling immunofluorescence studies with glomerular cell markers demonstrated that SV2B is expressed in glomerular visceral epithelial cells (podocytes). The expression of SV2B is detected also in cultured podocyte and in human kidney section as podocytic pattern. The decrease of SV2B mRNA was already detected before the onset of proteinuria in PAN nephropathy. The mRNA expression of SV2B clearly is altered not only in PAN nephropathy but also in another proteinuric state that is caused by an antibody against nephrin, a functional molecule of the slit diaphragm. The decreased intensity in SV2B staining was already detected before the peak of proteinuria in both models with immunofluorescence study. A reduced amount of SV2B was detected in both models also with Western blot analysis. CD2AP, another functional molecule of the slit diaphragm, was observed in cytoplasm, including the processes area of the cultured podocyte, and when the podocyte was treated with small interfering RNA for SV2B, CD2AP staining at the process area was not detected. These results suggest that SV2B is a functional molecule of podocyte, and SV2B may play a role in the expression and proper localization of CD2AP.
Insights
Synaptic vesicle protein 2B (SV2B) is decreased in proteinuric kidney disease before symptoms appear. SV2B appears crucial for podocyte function and the localization of CD2AP, a slit diaphragm protein.
Area of Science:
- Nephrology
- Molecular Biology
- Cell Biology
Background:
- Synaptic vesicle protein 2B (SV2B) is a protein expressed in podocytes.
- Reduced SV2B mRNA is observed in puromycin aminonucleoside (PAN) nephropathy.
- The role of SV2B in kidney disease is not well understood.
Purpose of the Study:
- To investigate the role of SV2B in podocyte function and proteinuric kidney disease.
- To determine if SV2B expression is altered in different models of kidney injury.
- To explore the relationship between SV2B and CD2AP in podocytes.
Main Methods:
- Glomerular cDNA subtraction assay to identify differentially expressed genes.
- Western blot analysis to detect SV2B protein levels.
- Dual-labeling immunofluorescence to study SV2B and CD2AP localization in podocytes and kidney sections.
Main Results:
- SV2B mRNA expression decreased in PAN nephropathy prior to proteinuria onset.
- SV2B expression was also reduced in an antibody-induced nephrin model of proteinuric kidney disease.
- SV2B staining intensity decreased before peak proteinuria in both models.
- Reduced SV2B levels were confirmed by Western blot.
- CD2AP localization to podocyte processes was lost upon SV2B knockdown, suggesting SV2B's role in CD2AP regulation.
Conclusions:
- SV2B is a functional molecule in podocytes.
- SV2B expression is altered in proteinuric kidney disease.
- SV2B may play a critical role in the expression and proper localization of CD2AP, a key slit diaphragm protein.
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