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 Ahn, Kenneth R. Newton, Armando J Mendez, Patricia I. Arnold, Duane R. Schultz

Research output: Contribution to journalArticle

21 Citations (Scopus)

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

Fingerprint

Mitomycin
Endothelial Cells
Tumor Necrosis Factor-alpha
Kidney
Lymphotoxin-beta
Proteins
Caspase 3
Lymphokines
Fluorescence Microscopy
Flow Cytometry
Theoretical Models
Membranes

Keywords

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

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

Cite this

Agonist-induced capping of adhesion proteins and microparticle shedding in cultures of human renal microvascular endothelial cells. / Jy, Wenche; Jimenez, Joaquin J; Mauro, Lucia M.; Ahn, Yeon; Newton, Kenneth R.; Mendez, Armando J; Arnold, Patricia I.; Schultz, Duane R.

In: Endothelium: Journal of Endothelial Cell Research, Vol. 9, No. 3, 24.09.2002, p. 179-189.

Research output: Contribution to journalArticle

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