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Isolation of Glomeruli and In Vivo Labeling of Glomerular Cell Surface Proteins
Published on: January 18, 2019
Phenotypic expression of ADAMTS13 in glomerular endothelial cells
Ramesh Tati1, Ann-Charlotte Kristoffersson, Anne-Lie Ståhl
1Department of Pediatrics, Clinical Sciences Lund, Lund University, Lund, Sweden.
Plos One
|July 2, 2011
Summary
Glomerular endothelial cells express ADAMTS13, a protease that cleaves von Willebrand factor. This activity may prevent platelet deposition in kidney capillaries under high shear stress.
Area of Science:
- Renal physiology
- Vascular biology
- Protease function
Background:
- ADAMTS13 is the key enzyme cleaving von Willebrand factor (VWF).
- Understanding ADAMTS13 expression and function in the kidney is crucial for renal health.
Purpose of the Study:
- To investigate ADAMTS13 expression and bioactivity in mouse and human kidney cells.
- To determine the role of ADAMTS13 in preventing platelet aggregation in glomerular capillaries.
Main Methods:
- Immunohistochemistry on mouse kidney sections.
- Ultramorphology for phenotypic analysis.
- Real-time PCR, flow cytometry, immunofluorescence, and immunoblotting in human endothelial cells.
- VWF multimer analysis and immunoblotting for cleavage activity.
Main Results:
- ADAMTS13 was detected in glomerular endothelial cells of wild-type mice but absent in deficient mice.
- ADAMTS13-deficient mice showed thickened glomerular capillaries and platelet deposition.
- Human glomerular and dermal microvascular endothelial cells express and secrete ADAMTS13.
- ADAMTS13 demonstrated VWF-cleaving activity in glomerular endothelial cells.
Conclusions:
- Glomerular endothelial cells are a source of ADAMTS13.
- ADAMTS13 secreted by endothelial cells likely protects glomerular capillaries from platelet deposition.
- This proteolytic activity is important in high shear stress environments like the glomerulus.
