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University of Vermont College of Medicine

4 ARTICLES PUBLISHED IN JoVE

JoVE Core

Measurement of Smooth Muscle Function in the Isolated Tissue Bath-applications to Pharmacology Research

Brian Jespersen¹, Nathan R. Tykocki², Stephanie W. Watts¹, Peter J. Cobbett¹

¹Department of Pharmacology and Toxicology, Michigan State University, ²Department of Pharmacology, University of Vermont College of Medicine

This protocol describes the measurement of isometric contraction in an isolated smooth muscle preparation, using an isolated tissue bath system and computer-based data acquisition.

Neuroscience

Isolation and Cannulation of Cerebral Parenchymal Arterioles

Paulo W. Pires¹, Fabrice Dabertrand², Scott Earley¹

¹Department of Pharmacology, University of Nevada School of Medicine, ²Department of Pharmacology, University of Vermont College of Medicine

This manuscript describes a simple and reproducible protocol for isolation of intracerebral arterioles (a group of blood vessels encompassing parenchymal arterioles, penetrating arterioles and pre-capillary arterioles) from mice, to be used in pressure myography, immunofluorescence, biochemistry, and molecular studies.

Neuroscience

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

Amanda C. Rosehart¹, Abbie C. Johnson², Fabrice Dabertrand^1,3

¹Department of Anesthesiology, University of Colorado Anschutz Medical Campus, ²Department of Neurological Sciences, University of Vermont Larner College of Medicine, ³Department of Pharmacology, University of Colorado Anschutz Medical Campus

The present manuscript details how to isolate hippocampal arterioles and capillaries from the mouse brain and how to pressurize them for pressure myography, immunofluorescence, biochemistry, and molecular studies.

Biochemistry

In Vitro Characterization of Histone Chaperones using Analytical, Pull-Down and Chaperoning Assays

Ruchir C. Bobde^*1,2, Ketul Saharan^*1,2, Somanath Baral^1,3, Surajit Gandhi^1,2, Archana Samal^1,2, Rajivgandhi Sundaram¹, Ashish Kumar^1,4, Ajit K. Singh^1,5, Aritreyee Datta¹, Dileep Vasudevan¹

¹Institute of Life Sciences, ²Regional Centre for Biotechnology, ³School of Biotechnology, KIIT University, ⁴Department of Molecular Biophysics and Biochemistry, Yale University, ⁵Department of Pharmacology, University of Vermont College of Medicine

This protocol describes a battery of methods that includes analytical size-exclusion chromatography to study histone chaperone oligomerization and stability, pull-down assay to unravel histone chaperone-histone interactions, AUC to analyze the stoichiometry of the protein complexes, and histone chaperoning assay to functionally characterize a putative histone chaperone in vitro.