Phenotypic expression of ADAMTS13 in glomerular endothelial cells

PLoS One. 2011;6(6):e21587. doi: 10.1371/journal.pone.0021587. Epub 2011 Jun 24.

Abstract

Background: ADAMTS13 is the physiological von Willebrand factor (VWF)-cleaving protease. The aim of this study was to examine ADAMTS13 expression in kidneys from ADAMTS13 wild-type (Adamts13⁺/⁺) and deficient (Adamts13⁻/⁻) mice and to investigate the expression pattern and bioactivity in human glomerular endothelial cells.

Methodology/principal findings: Immunohistochemistry was performed on kidney sections from ADAMTS13 wild-type and ADAMTS13-deficient mice. Phenotypic differences were examined by ultramorphology. ADAMTS13 expression in human glomerular endothelial cells and dermal microvascular endothelial cells was investigated by real-time PCR, flow cytometry, immunofluorescence and immunoblotting. VWF cleavage was demonstrated by multimer structure analysis and immunoblotting. ADAMTS13 was demonstrated in glomerular endothelial cells in Adamts13⁺/⁺ mice but no staining was visible in tissue from Adamts13⁻/⁻ mice. Thickening of glomerular capillaries with platelet deposition on the vessel wall was detected in Adamts13⁻/⁻ mice. ADAMTS13 mRNA and protein were detected in both human endothelial cells and the protease was secreted. ADAMTS13 activity was demonstrated in glomerular endothelial cells as cleavage of VWF.

Conclusions/significance: Glomerular endothelial cells express and secrete ADAMTS13. The proteolytic activity could have a protective effect preventing deposition of platelets along capillary lumina under the conditions of high shear stress present in glomerular capillaries.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAM Proteins / metabolism*
  • ADAMTS13 Protein
  • Animals
  • Blood Platelets / metabolism
  • Capillaries / metabolism
  • Cells, Cultured
  • Culture Media / metabolism
  • Endothelial Cells / metabolism*
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Gene Expression Regulation
  • Humans
  • Kidney Glomerulus / blood supply
  • Kidney Glomerulus / cytology*
  • Kidney Glomerulus / metabolism*
  • Kidney Glomerulus / ultrastructure
  • Matrix Metalloproteinase 9 / metabolism
  • Metalloendopeptidases / deficiency
  • Metalloendopeptidases / genetics
  • Metalloendopeptidases / metabolism*
  • Mice
  • Phenotype
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Substrate Specificity
  • von Willebrand Factor / chemistry
  • von Willebrand Factor / metabolism

Substances

  • Culture Media
  • RNA, Messenger
  • von Willebrand Factor
  • ADAM Proteins
  • ADAMTS13 protein, mouse
  • Metalloendopeptidases
  • Matrix Metalloproteinase 9
  • ADAMTS13 Protein
  • ADAMTS13 protein, human