title: Mitochondrial fission in endothelial cells after simulated ischemia/reperfusion: role of nitric oxide and reactive oxygen species.
abstract: Ischemia (I)/reperfusion (RP)-induced endothelial cell (EC) injury is thought to be due to mitochondrial reactive oxygen species (mtROS) production. MtROS have been implicated in mitochondrial fission. We determined whether cultured EC exposure to simulated I/RP causes morphological changes in the mitochondrial network and the mechanisms behind those changes. Because shear stress results in nitric oxide (NO)-mediated endothelial mtROS generation, we simulated I/RP as hypoxia (H) followed by oxygenated flow over the ECs (shear stress of 10dyn/cm(2)). By exposing ECs to shear stress, H, H/reoxygenation (RO), or simulated I/RP and employing MitoTracker staining, we assessed the differential effects of changes in mechanical forces and/or O(2) levels on the mitochondrial network. Static or sheared ECs maintained their mitochondrial network. H- or H/RO-exposed ECs underwent changes, but mitochondrial fission was significantly less compared to that in ECs exposed to I/RP. I/RP-induced fission was partially inhibited by antioxidants, a NO synthase inhibitor, or an inhibitor of the fission protein dynamin-related protein 1 (Drp1) and was accompanied by Drp1 oligomerization and phosphorylation (Ser616). Hence, shear-induced NO, ROS (including mtROS), and Drp1 activation are responsible for mitochondrial fission in I/RP-exposed ECs, and excessive fission may be an underlying cause of EC dysfunction in postischemic hearts.
authors.affiliation: Department of Biomedical Engineering, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Randy J
authors.initials: RJ
authors.lastname: Giedt
authors.forename: Changjun
authors.initials: C
authors.lastname: Yang
authors.forename: Jay L
authors.initials: JL
authors.lastname: Zweier
authors.forename: Anastasios
authors.initials: A
authors.lastname: Matzavinos
authors.forename: B Rita
authors.initials: BR
authors.lastname: Alevriadou
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D015687:Cell Hypoxia; D002478:Cells, Cultured; D034281:Dynamins; D020558:GTP Phosphohydrolases / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D053078:Membrane Potential, Mitochondrial; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000378:metabolism; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism; D053140:Organelle Shape; D062907:Oxaloacetic Acid / Q000494:pharmacology; D010766:Phosphorylation; D055503:Protein Multimerization; D015427:Reperfusion Injury / Q000378:metabolism*; D033081:Shear Strength; D013314:Stress, Mechanical; D013481:Superoxides / Q000378:metabolism*
chemical_list: D000975:Antioxidants; D008869:Microtubule-Associated Proteins; D024101:Mitochondrial Proteins; D013481:Superoxides; D062907:Oxaloacetic Acid; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D020558:GTP Phosphohydrolases; C110768:DNM1L protein, human; D034281:Dynamins; D019331:NG-Nitroarginine Methyl Ester; D000111:Acetylcysteine
references.citation: Mol Biol Cell. 2001 Aug;12(8):2245-56
references.pmid: 11514614
references.citation: J Alzheimers Dis. 2010;20 Suppl 2:S513-26
references.pmid: 20463395
references.citation: Biochem Biophys Res Commun. 2009 Oct 16;388(2):406-12
references.pmid: 19666009
references.citation: Nat Rev Mol Cell Biol. 2005 Aug;6(8):657-63
references.pmid: 16025099
references.citation: FEBS Lett. 1994 May 23;345(1):50-4
references.pmid: 8194600
references.citation: Circulation. 2010 May 11;121(18):2012-22
references.pmid: 20421521
references.citation: Circulation. 1995 Dec 15;92(12):3505-12
references.pmid: 8521573
references.citation: J Biol Chem. 2005 Jul 15;280(28):26185-92
references.pmid: 15899901
references.citation: Dev Cell. 2008 Feb;14(2):193-204
references.pmid: 18267088
references.citation: J Cell Biol. 2002 Dec 23;159(6):931-8
references.pmid: 12499352
references.citation: J Biol Chem. 1975 Sep 25;250(18):7114-9
references.pmid: 240815
references.citation: Am J Physiol Cell Physiol. 2008 Jul;295(1):C180-91
references.pmid: 18480296
references.citation: Annu Rev Physiol. 1995;57:311-32
references.pmid: 7778871
references.citation: Biochem Biophys Res Commun. 2000 Apr 13;270(2):616-21
references.pmid: 10753672
references.citation: EMBO J. 2006 Aug 9;25(15):3618-26
references.pmid: 16874301
references.citation: Dev Cell. 2001 Oct;1(4):515-25
references.pmid: 11703942
references.citation: Cardiovasc Res. 2010 Oct 1;88(1):16-29
references.pmid: 20631158
references.citation: Proc Natl Acad Sci U S A. 2011 Jun 21;108(25):10190-5
references.pmid: 21646527
references.citation: Biochim Biophys Acta. 2009 Jan;1793(1):154-70
references.pmid: 18694785
references.citation: Biophys J. 1999 Jan;76(1 Pt 1):469-77
references.pmid: 9876159
references.citation: J Pathol. 2000 Feb;190(3):255-66
references.pmid: 10685060
references.citation: PLoS One. 2009;4(5):e5546
references.pmid: 19436752
references.citation: PLoS Genet. 2010 Jan;6(1):e1000795
references.pmid: 20062521
references.citation: Am J Physiol Regul Integr Comp Physiol. 2004 Nov;287(5):R1014-30
references.pmid: 15475499
references.citation: Ann Biomed Eng. 2007 May;35(5):683-93
references.pmid: 17340195
references.citation: Br J Pharmacol. 2006 Jan;147 Suppl 1:S193-201
references.pmid: 16402104
references.citation: Am J Physiol Cell Physiol. 2008 Jun;294(6):C1576-85
references.pmid: 18434620
references.citation: Cardiovasc Res. 2006 May 1;70(2):181-90
references.pmid: 16580655
references.citation: Semin Cell Dev Biol. 2010 Aug;21(6):558-65
references.pmid: 20025987
references.citation: Proc Natl Acad Sci U S A. 2006 Feb 21;103(8):2653-8
references.pmid: 16477035
references.citation: Cardiovasc Res. 2004 Feb 15;61(3):402-13
references.pmid: 14962472
references.citation: Biochim Biophys Acta. 2010 Jan;1802(1):162-6
references.pmid: 19747972
references.citation: Biochemistry. 2008 Jul 8;47(27):7256-63
references.pmid: 18553936
references.citation: Proc Natl Acad Sci U S A. 2008 Oct 14;105(41):15803-8
references.pmid: 18838687
references.citation: Circ Res. 2002 Apr 5;90(6):745-8
references.pmid: 11934844
references.citation: Circ Res. 1999 Apr 16;84(7):840-5
references.pmid: 10205152
references.citation: Science. 2004 Sep 17;305(5691):1747-52
references.pmid: 15297626
references.citation: Br J Pharmacol. 2003 Feb;138(4):532-43
references.pmid: 12598407
references.citation: Circulation. 1990 Oct;82(4):1402-12
references.pmid: 2401073
references.citation: Cardiovasc Res. 2008 Jan 15;77(2):334-43
references.pmid: 18006487
references.citation: Am J Physiol Regul Integr Comp Physiol. 2004 Mar;286(3):R431-44
references.pmid: 14761864
references.citation: EMBO Rep. 2007 Oct;8(10):939-44
references.pmid: 17721437
references.citation: Arch Biochem Biophys. 1996 Apr 1;328(1):85-92
references.pmid: 8638942
references.citation: Biochim Biophys Acta. 2011 May;1807(5):491-502
references.pmid: 21406178
references.citation: Circulation. 2005 Jun 7;111(22):2966-72
references.pmid: 15939832
references.citation: J Clin Invest. 2005 Oct;115(10):2640-7
references.pmid: 16200197
references.citation: Proc Natl Acad Sci U S A. 2007 Sep 18;104(38):15081-6
references.pmid: 17848522
references.citation: Proc Natl Acad Sci U S A. 1999 Feb 16;96(4):1559-62
references.pmid: 9990063
references.citation: PLoS One. 2009 May 27;4(5):e5701
references.pmid: 19492085
references.citation: J Pharmacol Exp Ther. 2009 Apr;329(1):94-101
references.pmid: 19131585
references.citation: Science. 2009 Apr 3;324(5923):102-5
references.pmid: 19342591
references.citation: Curr Biol. 2006 Jul 25;16(14):R551-60
references.pmid: 16860735
references.citation: Hum Mol Genet. 2005 Jun 1;14(11):1405-15
references.pmid: 15829499
references.citation: J Cell Sci. 2007 Mar 1;120(Pt 5):838-48
references.pmid: 17298981
references.citation: Curr Mol Med. 2003 Sep;3(6):527-35
references.pmid: 14527084
references.citation: Antioxid Redox Signal. 2011 Sep 15;15(6):1517-30
references.pmid: 21194353
references.citation: Curr Biol. 2004 Feb 17;14(4):340-5
references.pmid: 14972687
references.citation: J Biol Chem. 1999 Dec 31;274(53):37709-16
references.pmid: 10608829
references.citation: J Biol Chem. 2007 Apr 13;282(15):11521-9
references.pmid: 17301055
references.citation: Am J Physiol Cell Physiol. 2007 Mar;292(3):C1103-12
references.pmid: 17020931
references.citation: J Cell Biol. 2008 Aug 11;182(3):573-85
references.pmid: 18695047
references.citation: Biotechniques. 2007 Jul;43(1 Suppl):25-30
references.pmid: 17936939
references.citation: EMBO J. 2006 Jul 12;25(13):2966-77
references.pmid: 16778770
references.citation: Science. 1985 Mar 22;227(4693):1477-9
references.pmid: 3883488
references.citation: J Biol Chem. 1998 Oct 2;273(40):25804-8
references.pmid: 9748253
references.citation: Circ Res. 2007 Apr 27;100(8):1128-41
references.pmid: 17463328
references.citation: Biochim Biophys Acta. 2006 May-Jun;1757(5-6):518-24
references.pmid: 16829229
references.citation: J Biol Chem. 2006 Dec 8;281(49):37972-9
references.pmid: 17003040
references.citation: Cytometry A. 2006 Jan;69(1):1-12
references.pmid: 16342116
references.citation: J Biol Chem. 2007 Aug 17;282(33):23788-98
references.pmid: 17537722
references.citation: Proc Natl Acad Sci U S A. 2006 Apr 4;103(14):5379-84
references.pmid: 16565215