title: Stepwise recruitment of transcellular and paracellular pathways underlies blood-brain barrier breakdown in stroke.
abstract: Brain endothelial cells form a paracellular and transcellular barrier to many blood-borne solutes via tight junctions (TJs) and scarce endocytotic vesicles. The blood-brain barrier (BBB) plays a pivotal role in the healthy and diseased CNS. BBB damage after ischemic stroke contributes to increased mortality, yet the contributions of paracellular and transcellular mechanisms to this process in vivo are unknown. We have created a transgenic mouse strain whose endothelial TJs are labeled with eGFP and have imaged dynamic TJ changes and fluorescent tracer leakage across the BBB in vivo, using two-photon microscopy in the t-MCAO stroke model. Although barrier function is impaired as early as 6 hr after stroke, TJs display profound structural defects only after 2 days. Conversely, the number of endothelial caveolae and transcytosis rate increase as early as 6 hr after stroke. Therefore, stepwise impairment of transcellular followed by paracellular barrier mechanisms accounts for the BBB deficits in stroke.
authors.affiliation: Department of Developmental and Cell Biology, University of California, Irvine, CA 92697, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Knowland
authors.affiliation: Department of Neurosurgery, Stanford University School of Medicine, Stanford, CA 94305, USA; Department of Medicine, University of California, Irvine, Orange, CA 92868, USA.
authors.forename: Ahmet
authors.initials: A
authors.lastname: Arac
authors.affiliation: Waitt Advanced Biophotonics Center, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA; Biological Sciences Graduate Program, University of California, San Diego, La Jolla, CA 92037, USA.
authors.forename: Kohei J
authors.initials: KJ
authors.lastname: Sekiguchi
authors.affiliation: Department of Developmental and Cell Biology, University of California, Irvine, CA 92697, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Hsu
authors.affiliation: Department of Developmental and Cell Biology, University of California, Irvine, CA 92697, USA.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Lutz
authors.affiliation: Electron Microscopy Facility, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: John
authors.initials: J
authors.lastname: Perrino
authors.affiliation: Department of Neurosurgery, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Gary K
authors.initials: GK
authors.lastname: Steinberg
authors.affiliation: Department of Neurobiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Ben A
authors.initials: BA
authors.lastname: Barres
authors.affiliation: Waitt Advanced Biophotonics Center, The Salk Institute for Biological Studies, La Jolla, CA 92037, USA. Electronic address: animmerj@salk.edu.
authors.forename: Axel
authors.initials: A
authors.lastname: Nimmerjahn
authors.affiliation: Department of Developmental and Cell Biology, University of California, Irvine, CA 92697, USA. Electronic address: dagalliu@uci.edu.
authors.forename: Dritan
authors.initials: D
authors.lastname: Agalliu
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000378:metabolism* / Q000473:pathology* / Q000648:ultrastructure; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D020521:Stroke / Q000378:metabolism* / Q000473:pathology*; D019108:Tight Junctions / Q000378:metabolism / Q000473:pathology; D057900:Transcytosis / Q000502:physiology*
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