Oxidative stress increases blood-brain barrier permeability and induces alterations in occludin during hypoxia-reoxygenation.
The blood-brain barrier (BBB) has a critical role in central nervous system homeostasis. Intercellular tight junction (TJ) protein complexes of the brain microvasculature limit paracellular diffusion of substances from the blood into the brain. Hypoxia and reoxygenation (HR) is a central component to numerous disease states and pathologic conditions. We have previously shown that HR can influence the permeability of the BBB as well as the critical TJ protein occludin. During HR, free radicals are produced, which may lead to oxidative stress. Using the free radical scavenger tempol (200 mg/kg, intraperitoneal), we show that oxidative stress produced during HR (6% O(2) for 1 h, followed by room air for 20 min) mediates an increase in BBB permeability in vivo using in situ brain perfusion. We also show that these changes are associated with alterations in the structure and localization of occludin. Our data indicate that oxidative stress is associated with movement of occludin away from the TJ. Furthermore, subcellular fractionation of cerebral microvessels reveals alterations in occludin oligomeric assemblies in TJ associated with plasma membrane lipid rafts. Our data suggest that pharmacological inhibition of disease states with an HR component may help preserve BBB functional integrity.
Program in Neuroscience, University of Arizona College of Medicine, Tucson, Arizona 85724, USA.
Jeffrey J
JJ
Lochhead
Gwen
G
McCaffrey
Colleen E
CE
Quigley
Jessica
J
Finch
Kristin M
KM
DeMarco
Nicole
N
Nametz
Thomas P
TP
Davis
D000818:Animals; D001812:Blood-Brain Barrier / Q000502:physiology*; D015153:Blotting, Western; D002196:Capillaries / Q000378:metabolism / Q000502:physiology; D002499:Centrifugation, Density Gradient; D002560:Cerebrovascular Circulation / Q000502:physiology; D003497:Cyclic N-Oxides / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005455:Fluorescent Antibody Technique; D016166:Free Radical Scavengers / Q000494:pharmacology; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis; D002534:Hypoxia, Brain / Q000378:metabolism* / Q000473:pathology; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000096:biosynthesis; D007202:Indicators and Reagents; D008565:Membrane Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal; D062793:Occludin; D018384:Oxidative Stress / Q000502:physiology*; D010539:Permeability; D051381:Rats; D017207:Rats, Sprague-Dawley; D013113:Spin Labels; D014178:Translocation, Genetic
D003497:Cyclic N-Oxides; D016166:Free Radical Scavengers; D018840:HSP70 Heat-Shock Proteins; C497444:Hif1a protein, rat; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D007202:Indicators and Reagents; D008565:Membrane Proteins; D062793:Occludin; C568987:Ocln protein, rat; D013113:Spin Labels; C001803:tempol