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      Ex Vivo Pressurized Hippocampal Capillary-Parenchymal Arteriole Preparation for Functional Study

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          Abstract

          From subtle behavioral alterations to late-stage dementia, vascular cognitive impairment typically develops following cerebral ischemia. Stroke and cardiac arrest are remarkably sexually dimorphic diseases, and both induce cerebral ischemia. However, progress in understanding the vascular cognitive impairment, and then developing sex-specific treatments, has been partly limited by challenges in investigating the brain microcirculation from mouse models in functional studies. Here, we present an approach to examine the capillary-to-arteriole signaling in an ex vivo hippocampal capillary-parenchymal arteriole (HiCaPA) preparation from mouse brain. We describe how to isolate, cannulate, and pressurize the microcirculation to measure arteriolar diameter in response to capillary stimulation. We show which appropriate functional controls can be used to validate the HiCaPA preparation integrity and display typical results, including testing potassium as a neurovascular coupling agent and the effect of the recently characterized inhibitor of the Kir2 inward rectifying potassium channel family, ML133. Further, we compare the responses in preparations obtained from male and female mice. While these data reflect functional investigations, our approach can also be used in molecular biology, immunochemistry, and electrophysiology studies.

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          Author and article information

          Journal
          101313252
          34584
          J Vis Exp
          J Vis Exp
          Journal of visualized experiments : JoVE
          1940-087X
          13 January 2020
          18 December 2019
          18 December 2019
          29 January 2020
          : 154
          : 10.3791/60676
          Affiliations
          [1 ]Department of Anesthesiology, University of Colorado Anschutz Medical Campus
          [2 ]Department of Neurological Sciences, University of Vermont Larner College of Medicine
          [3 ]Department of Pharmacology, University of Colorado Anschutz Medical Campus
          Author notes
          Correspondence to: Fabrice Dabertrand at Fabrice.Dabertrand@ 123456CUAnschutz.edu
          Article
          PMC6988500 PMC6988500 6988500 nihpa1067401
          10.3791/60676
          6988500
          31904015
          0926bd87-4e6e-49c8-888e-003f4771d471
          History
          Categories
          Article

          microcirculation,neurovascular coupling,Neuroscience,pressure myography,hippocampus,ion channel,cerebral parenchymal arterioles,myogenic reactivity,brain capillary,capillary to arteriole electrical signaling,Issue 154,vessel cannulation

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