Journal article
Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice
Journal of Visualized Experiments, Vol.2015(99), pp.e52675-e52675
2015
DOI: 10.3791/52675
PMCID: PMC4541629
PMID: 25993394
Abstract
Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week protocol for repetitive hypoxic preconditioning (RHP) induces long-term protection against central nervous system (CNS) injury in a mouse model of focal ischemic stroke. RHP consists of 9 stochastic exposures to hypoxia that vary in both duration (2 or 4 hr) and intensity (8% and 11% O2). RHP reduces infarct volumes, blood-brain barrier (BBB) disruption, and the post-stroke inflammatory response for weeks following the last exposure to hypoxia, suggesting a long-term induction of an endogenous CNS-protective phenotype. The methodology for the dual quantification of infarct volume and BBB disruption is effective in assessing neurovascular protection in mice with RHP or other putative neuroprotectants. Adult male Swiss Webster mice were preconditioned by RHP or duration-equivalent exposures to 21% O2 (i.e. room air). A 60 min transient middle cerebral artery occlusion (tMCAo) was induced 2 weeks following the last hypoxic exposure. Both the occlusion and reperfusion were confirmed by transcranial laser Doppler flowmetry. Twenty-two hr after reperfusion, Evans Blue (EB) was intravenously administered through a tail vein injection. 2 hr later, animals were sacrificed by isoflurane overdose and brain sections were stained with 2,3,5- triphenyltetrazolium chloride (TTC). Infarcts volumes were then quantified. Next, EB was extracted from the tissue over 48 hr to determine BBB disruption after tMCAo. In summary, RHP is a simple protocol that can be replicated, with minimal cost, to induce long-term endogenous neurovascular protection from stroke injury in mice, with the translational potential for other CNS-based and systemic pro-inflammatory disease states.
Details
- Title: Subtitle
- Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice
- Creators
- Katherine Poinsatte - Department of Neurology and Neurotherapeutics, University of Texas Southwestern Medical CenterUma Maheswari Selvaraj - Department of Neurology and Neurotherapeutics, University of Texas Southwestern Medical CenterSterling B Ortega - Department of Neurology and Neurotherapeutics, University of Texas Southwestern Medical CenterErik J Plautz - Department of Neurology and Neurotherapeutics, University of Texas Southwestern Medical CenterXiangmei Kong - Department of Neurology and Neurotherapeutics, University of Texas Southwestern Medical CenterJeffrey M Gidday - Department of Neurological Surgery, Washington University School of MedicineAnn M Stowe - Department of Neurology and Neurotherapeutics, University of Texas Southwestern Medical Center
- Resource Type
- Journal article
- Publication Details
- Journal of Visualized Experiments, Vol.2015(99), pp.e52675-e52675
- Publisher
- MyJove Corporation
- DOI
- 10.3791/52675
- PMID
- 25993394
- PMCID
- PMC4541629
- ISSN
- 1940-087X
- eISSN
- 1940-087X
- Language
- English
- Date published
- 2015
- Academic Unit
- Pathology
- Record Identifier
- 9984065385502771
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