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1 Department of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ, USA
* To whom correspondence should be addressed. E-mail: kawitt{at}u.arizona.edu.
Cerebral microvessel endothelial cells that form the blood-brain barrier (BBB) have tight junctions (TJ) that are critical for maintaining brain homeostasis. The effects of initial reoxygenation following a hypoxic insult (H/R) on functional and molecular properties of the BBB and TJ remain unclear. In situ brain perfusion and Western blot analyses were performed to assess in vivo BBB integrity upon reoxygenation, following a hypoxic insult of 6% O2/ 1 hr. Model conditions (blood-pressure, blood-gas chemistries, cerebral-blood flow, and brain ATP concentration), to ensure a consistent level and criteria for insult, were also assessed. In situ brain perfusion determined that initial reoxygenation (10 min) significantly increased the uptake of [14C] sucrose into brain parenchyma. Capillary depletion and cerebral blood-flow analyses indicate the perturbations were due to increased paracellular permeability rather than vascular volume changes. Hypoxia with reoxygenation (10 min) produced an increase in BBB permeability with associated alterations in TJ protein expression. These results suggest that H/R leads to reorganization of TJs and increased paracellular diffusion at the BBB, which is not a result of increased CBF, vascular volume change, or endothelial uptake of marker. Additionally, the TJ protein occludin had a shift in bands correlated with functional changes (i.e increased permeability), without significant change in expression of claudin-3, ZO-1, or actin. H/R induced changes in the BBB may result in edema and/or associated pathological outcomes.
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