title: Rapid endothelial cytoskeletal reorganization enables early blood-brain barrier disruption and long-term ischaemic reperfusion brain injury.
abstract: The mechanism and long-term consequences of early blood-brain barrier (BBB) disruption after cerebral ischaemic/reperfusion (I/R) injury are poorly understood. Here we discover that I/R induces subtle BBB leakage within 30-60 min, likely independent of gelatinase B/MMP-9 activities. The early BBB disruption is caused by the activation of ROCK/MLC signalling, persistent actin polymerization and the disassembly of junctional proteins within microvascular endothelial cells (ECs). Furthermore, the EC alterations facilitate subsequent infiltration of peripheral immune cells, including MMP-9-producing neutrophils/macrophages, resulting in late-onset, irreversible BBB damage. Inactivation of actin depolymerizing factor (ADF) causes sustained actin polymerization in ECs, whereas EC-targeted overexpression of constitutively active mutant ADF reduces actin polymerization and junctional protein disassembly, attenuates both early- and late-onset BBB impairment, and improves long-term histological and neurological outcomes. Thus, we identify a previously unexplored role for early BBB disruption in stroke outcomes, whereby BBB rupture may be a cause rather than a consequence of parenchymal cell injury.
authors.affiliation: Center of Cerebrovascular Disease Research, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.|China-America Institute of Neuroscience, Xuanwu Hospital, Capital Medical University, Beijing 100053, China.
authors.forename: Yejie
authors.identifier: http://orcid.org/0000-0001-7502-9201
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Center of Cerebrovascular Disease Research, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.|State Key Laboratory of Medical Neurobiology, Institute of Brain Science and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China.|Geriatric Research, Educational and Clinical Center, Veterans Affairs Pittsburgh Health Care System, Pittsburgh, Pennsylvania 15261, USA.
authors.forename: Lili
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Center of Cerebrovascular Disease Research, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.|State Key Laboratory of Medical Neurobiology, Institute of Brain Science and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China.
authors.forename: Hongjian
authors.initials: H
authors.lastname: Pu
authors.affiliation: Center of Cerebrovascular Disease Research, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.|State Key Laboratory of Medical Neurobiology, Institute of Brain Science and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China.
authors.forename: Leilei
authors.initials: L
authors.lastname: Mao
authors.affiliation: Center of Cerebrovascular Disease Research, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.|State Key Laboratory of Medical Neurobiology, Institute of Brain Science and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China.|Geriatric Research, Educational and Clinical Center, Veterans Affairs Pittsburgh Health Care System, Pittsburgh, Pennsylvania 15261, USA.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Hu
authors.affiliation: Center of Cerebrovascular Disease Research, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.|State Key Laboratory of Medical Neurobiology, Institute of Brain Science and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China.
authors.forename: Xiaoyan
authors.identifier: http://orcid.org/0000-0003-3832-3756
authors.initials: X
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Medical Neurobiology, Institute of Brain Science and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China.
authors.forename: Na
authors.initials: N
authors.lastname: Xu
authors.affiliation: Center of Cerebrovascular Disease Research, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.|State Key Laboratory of Medical Neurobiology, Institute of Brain Science and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China.|Geriatric Research, Educational and Clinical Center, Veterans Affairs Pittsburgh Health Care System, Pittsburgh, Pennsylvania 15261, USA.
authors.forename: R Anne
authors.initials: RA
authors.lastname: Stetler
authors.affiliation: Center of Cerebrovascular Disease Research, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.|State Key Laboratory of Medical Neurobiology, Institute of Brain Science and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China.|Geriatric Research, Educational and Clinical Center, Veterans Affairs Pittsburgh Health Care System, Pittsburgh, Pennsylvania 15261, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Center of Cerebrovascular Disease Research, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.|China-America Institute of Neuroscience, Xuanwu Hospital, Capital Medical University, Beijing 100053, China.
authors.forename: Xiangrong
authors.initials: X
authors.lastname: Liu
authors.affiliation: Division of Pharmaceutical Sciences, Mylan School of Pharmacy, Duquesne University, Pittsburgh, Pennsylvania 15282, USA.
authors.forename: Rehana K
authors.initials: RK
authors.lastname: Leak
authors.affiliation: Department of Neurosurgery, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Richard F
authors.initials: RF
authors.lastname: Keep
authors.affiliation: China-America Institute of Neuroscience, Xuanwu Hospital, Capital Medical University, Beijing 100053, China.
authors.forename: Xunming
authors.initials: X
authors.lastname: Ji
authors.affiliation: Center of Cerebrovascular Disease Research, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.|State Key Laboratory of Medical Neurobiology, Institute of Brain Science and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China.|Geriatric Research, Educational and Clinical Center, Veterans Affairs Pittsburgh Health Care System, Pittsburgh, Pennsylvania 15261, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Chen
mesh_terms: D000199:Actins / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001812:Blood-Brain Barrier / Q000378:metabolism*; D002545:Brain Ischemia / Q000235:genetics / Q000378:metabolism*; D003599:Cytoskeleton / Q000235:genetics / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D058105:Polymerization; D015427:Reperfusion Injury / Q000235:genetics / Q000378:metabolism*
chemical_list: D000199:Actins; D020780:Matrix Metalloproteinase 9
references.citation: J Cereb Blood Flow Metab. 2000 Oct;20(10):1474-82
references.pmid: 11043910
references.citation: BMC Neurosci. 2010;11:12
references.pmid: 20122276
references.citation: Front Neurosci. 2015 Apr 29;9:147
references.pmid: 25972779
references.citation: Cardiovasc Res. 2010 Jul 15;87(2):291-9
references.pmid: 20299333
references.citation: Circ Res. 2013 Sep 27;113(8):1013-22
references.pmid: 23780386
references.citation: Neuron. 2010 Jul 29;67(2):181-98
references.pmid: 20670828
references.citation: J Exp Med. 2001 Mar 19;193(6):741-54
references.pmid: 11257140
references.citation: J Clin Invest. 2013 Feb;123(2):782-6
references.pmid: 23281396
references.citation: J Exp Med. 2006 Apr 17;203(4):1007-19
references.pmid: 16585265
references.citation: J Cereb Blood Flow Metab. 2013 Jul;33(7):1104-14
references.pmid: 23571276
references.citation: J Neurosci. 2001 Oct 1;21(19):7724-32
references.pmid: 11567062
references.citation: Nat Neurosci. 2011 Nov;14(11):1363-8
references.pmid: 22030546
references.citation: Biophys J. 2004 Jun;86(6):3993-4003
references.pmid: 15189895
references.citation: Nat Methods. 2013 Mar;10(3):211-20
references.pmid: 23443633
references.citation: Cell Rep. 2015 Feb 24;10(7):1040-54
references.pmid: 25704809
references.citation: Microvasc Res. 2009 Jan;77(1):53-63
references.pmid: 19028505
references.citation: J Cereb Blood Flow Metab. 2006 Jun;26(6):797-810
references.pmid: 16192992
references.citation: J Biol Chem. 2009 Jul 10;284(28):19053-66
references.pmid: 19423710
references.citation: Curr Neuropharmacol. 2008 Sep;6(3):179-92
references.pmid: 19506719
references.citation: J Neurosci. 2008 Dec 3;28(49):13038-55
references.pmid: 19052195
references.citation: J Appl Physiol (1985). 2001 Oct;91(4):1487-500
references.pmid: 11568129
references.citation: Circ Res. 1982 Nov;51(5):657-61
references.pmid: 6890416
references.citation: Nat Med. 2011 Jul;17(7):796-808
references.pmid: 21738161
references.citation: J Cereb Blood Flow Metab. 2015 Jul;35(7):1191-8
references.pmid: 25757751
references.citation: Stroke. 2000 Aug;31(8):1965-72; discussion 1972-3
references.pmid: 10926965
references.citation: J Neurosci. 1998 Jul 1;18(13):4914-28
references.pmid: 9634557
references.citation: Biochim Biophys Acta. 2008 Mar;1778(3):794-809
references.pmid: 17961505
references.citation: Neurobiol Aging. 1988 Jan-Feb;9(1):31-9
references.pmid: 3288893
references.citation: J Immunol. 2014 Jan 1;192(1):324-37
references.pmid: 24259506
references.citation: Am J Physiol Heart Circ Physiol. 2005 Aug;289(2):H558-68
references.pmid: 15764676
references.citation: J Cereb Blood Flow Metab. 2003 Dec;23(12):1430-40
references.pmid: 14663338
references.citation: Integr Biol (Camb). 2009 Jun;1(5-6):404-26
references.pmid: 20023747
references.citation: Trends Cell Biol. 2002 Dec;12(12):598-605
references.pmid: 12495849
references.citation: Nature. 1984 Feb 16-22;307(5952):648-9
references.pmid: 6537993
references.citation: J Cereb Blood Flow Metab. 2011 Sep;31(9):1874-85
references.pmid: 21610723
references.citation: Ann Biomed Eng. 2010 Aug;38(8):2499-511
references.pmid: 20361260
references.citation: Lancet. 2000 May 13;355(9216):1670-4
references.pmid: 10905241
references.citation: J Neurosci. 2012 Feb 29;32(9):3044-57
references.pmid: 22378877
references.citation: J Cell Biol. 2013 Jan 7;200(1):9-19
references.pmid: 23295347
references.citation: Neuroscience. 2009 Feb 6;158(3):972-82
references.pmid: 18824084
references.citation: J Neurosci. 2010 May 5;30(18):6398-408
references.pmid: 20445066
references.citation: Stroke. 2009 Jun;40(6):2244-50
references.pmid: 19246690
references.citation: Nat Rev Neurosci. 2011 Mar;12(3):169-82
references.pmid: 21331083
references.citation: Neurology. 2012 Sep 25;79(13 Suppl 1):S52-7
references.pmid: 23008413
references.citation: Physiol Rev. 2006 Jan;86(1):279-367
references.pmid: 16371600
references.citation: Physiol Rev. 2003 Apr;83(2):433-73
references.pmid: 12663865
references.citation: J Neurosci. 2006 Mar 29;26(13):3491-5
references.pmid: 16571756
references.citation: J Cereb Blood Flow Metab. 2001 Apr;21(4):321-33
references.pmid: 11323518
references.citation: J Cereb Blood Flow Metab. 1999 Jun;19(6):624-33
references.pmid: 10366192
references.citation: Neuron. 2014 May 7;82(3):603-17
references.pmid: 24746419
references.citation: Am J Physiol Renal Physiol. 2006 Jun;290(6):F1398-407
references.pmid: 16434575
references.citation: Int J Stroke. 2009 Jun;4(3):159-68
references.pmid: 19659815
references.citation: Stroke. 2010 Sep;41(9):2056-63
references.pmid: 20651275
references.citation: J Cereb Blood Flow Metab. 2005 May;25(5):593-606
references.pmid: 15689955
references.citation: J Cereb Blood Flow Metab. 1998 Dec;18(12):1336-45
references.pmid: 9850146
references.citation: J Cereb Blood Flow Metab. 2000 Dec;20(12):1681-9
references.pmid: 11129784
references.citation: J Cereb Blood Flow Metab. 2007 Apr;27(4):697-709
references.pmid: 16850029
references.citation: Stroke. 2007 Apr;38(4):1345-53
references.pmid: 17332467
references.citation: Nat Med. 2006 Apr;12(4):441-5
references.pmid: 16565723
references.citation: Crit Rev Biochem Mol Biol. 2010 Oct;45(5):351-423
references.pmid: 20812779
references.citation: CNS Neurol Disord Drug Targets. 2013 May 1;12(3):325-37
references.pmid: 23469858
references.citation: Ann N Y Acad Sci. 2008 Mar;1123:134-45
references.pmid: 18375586
references.citation: J Cell Sci. 2007 Oct 15;120(Pt 20):3491-9
references.pmid: 17928305
references.citation: J Clin Invest. 1999 Feb;103(3):347-54
references.pmid: 9927495
references.citation: Ann Neurol. 2013 Sep;74(3):458-71
references.pmid: 23674483
references.citation: Neuroscience. 1998 Oct;86(4):1245-57
references.pmid: 9697130
references.citation: Mol Cell Biol. 2004 Jul;24(13):5937-52
references.pmid: 15199148