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4 ARTICLES PUBLISHED IN JoVE

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Biology

Visualization of MG53-mediated Cell Membrane Repair Using in vivo and in vitro Systems
Noah Weisleder 1, Peihui Lin 1, Xiaoli Zhao 1, Matthew Orange 1, Hua Zhu 1, Jianjie Ma 1
1Department of Physiology and Biophysics, Robert Wood Johnson Medical School

Described here are protocols used to visualize the dynamic process of MG53-mediated cell membrane repair in whole animals and at the cellular level. These methods can be applied to investigate the cell biology of plasma membrane resealing and regenerative medicine.

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Biology

Fluorescence-based Measurement of Store-operated Calcium Entry in Live Cells: from Cultured Cancer Cell to Skeletal Muscle Fiber
Zui Pan 1, Xiaoli Zhao 2, Marco Brotto 3
1Department of Physiology and Biophysics, Confocal Microscopy and Cell Imaging Core, Robert Wood Johnson Medical School , 2Department of Physiology and Biophysics, Robert Wood Johnson Medical School , 3Muscle Biology Research Group-MUBIG Schools of Nursing & Medicine, University of Missouri-Kansas City

The extent of store-operated Ca2+ entry (SOCE) can be monitored using fluorescent Ca2+ indicators. Mn2+ quenching of such indicators assays SOCE in cultured cells and skeletal muscle fibers. A technique allowing spatial and temporal resolution of SOCE by confocal imaging of mechanically skinned muscle fibers is also described.

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Biology

Ex Vivo Assessment of Contractility, Fatigability and Alternans in Isolated Skeletal Muscles
Ki Ho Park 1, Leticia Brotto 2, Oanh Lehoang 1, Marco Brotto 2, Jianjie Ma 1, Xiaoli Zhao 1,3
1Department of Physiology and Biophysics, UMDNJ-Robert Wood Johnson Medical School, 2Muscle Biology Research Group, University of Missouri-Kansas City, 3Pharmacology division, College of Pharmacy, DHLRI, Ohio State University

We describe a method to directly measure muscle force, muscle power, contractile kinetics and fatigability of isolated skeletal muscles in an in vitro system using field stimulation. Valuable information on Ca2+ handling properties and contractile machinery of the muscle can be obtained using different stimulating protocols.

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Immunology and Infection

Alveolar Macrophage Phagocytosis and Bacteria Clearance in Mice
Nagaraja Nagre 1, Xiaofei Cong 1, Andrew C. Pearson 1, Xiaoli Zhao 1
1Department of Physiological Sciences, Eastern Virginia Medical School

Here we report common methods to analyze the phagocytic function of murine alveolar macrophages and bacterial clearance from the lung. These methods study in vitro phagocytosis of fluorescein isothiocyanate beads and in vivo phagocytosis of Pseudomonas aeruginosa Green Fluorescent Protein. We also describe a method for clearing P. aeruginosa in mice.

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