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 language | English |
|---|---|
| Pages (from-to) | 179-189 |
| Number of pages | 11 |
| Journal | Endothelium: Journal of Endothelial Cell Research |
| Volume | 9 |
| Issue number | 3 |
| DOIs | |
| State | Published - Sep 24 2002 |
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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 journal › Article
}
TY - JOUR
T1 - Agonist-induced capping of adhesion proteins and microparticle shedding in cultures of human renal microvascular endothelial cells
AU - Jy, Wenche
AU - Jimenez, Joaquin J
AU - Mauro, Lucia M.
AU - Ahn, Yeon
AU - Newton, Kenneth R.
AU - Mendez, Armando J
AU - Arnold, Patricia I.
AU - Schultz, Duane R.
PY - 2002/9/24
Y1 - 2002/9/24
N2 - 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.
AB - 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.
KW - Caps
KW - Endothelial cells
KW - Immunofluorescence
KW - Microparticles
KW - Mitomycin C
KW - TNF-α
UR - http://www.scopus.com/inward/record.url?scp=0036040936&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0036040936&partnerID=8YFLogxK
U2 - 10.1080/10623320213632
DO - 10.1080/10623320213632
M3 - Article
C2 - 12380643
AN - SCOPUS:0036040936
VL - 9
SP - 179
EP - 189
JO - Endothelium: Journal of Endothelial Cell Research
JF - Endothelium: Journal of Endothelial Cell Research
SN - 1062-3329
IS - 3
ER -