Agonist-induced capping of adhesion proteins and microparticle shedding in cultures of human renal microvascular endothelial cells

Wenche Jy, Joaquin J. Jimenez, Lucia M. Mauro, Yeon S. Ahn, Kenneth R. Newton, Armando J. Mendez, Patricia I. Arnold, Duane R. Schultz

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

Capping and release of membranous, small (<1.5 μm) endothelial microparticles were quantified by immunofluorescence microscopy and flow cytometry after treatment of cultures of human renal microvascular endothelial cells with agonists tumor necrosis factor-alpha (TNF-α) or mitomycin C. For constitutive marker CD31, both agonist-treated attached, monolayer, and detached, free endothelial cells formed caps and released microparticles. TNF-α and mitomycin C induced dissimilar appearing CD31-containing caps after 3 h, followed by endothelial microparticle release after 6 h. The degree of capping correlated with increasing counts of released microparticles. For lymphokine-inducible CD54, TNF-α also induced CD54-containing caps and microparticle release, but mitomycin C failed to induce the expression of either entity. Neither capping nor microparticle release caused by TNF-α was part of an apoptotic pathway that involved caspase 3. Mitomycin C treatment of endothelial cells caused capping and microparticle release with a time course similar to TNF-α induction for 15 to 24 h, but assays for caspase 3 were positive, confirming the apoptotic action of mitomycin C. Membrane capping and microparticle release from endothelial cells are a convenient experimental model for studying protein movement, release of microparticles, and their possible biological significance.

Original languageEnglish (US)
Pages (from-to)179-189
Number of pages11
JournalEndothelium: Journal of Endothelial Cell Research
Volume9
Issue number3
DOIs
StatePublished - Sep 24 2002

Keywords

  • Caps
  • Endothelial cells
  • Immunofluorescence
  • Microparticles
  • Mitomycin C
  • TNF-α

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

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