S'identifier

University of Washington, Seattle

2 ARTICLES PUBLISHED IN JoVE

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Biology

Visualization of Vascular Ca2+ Signaling Triggered by Paracrine Derived ROS
Karthik Mallilankaraman 1, Rajesh Kumar Gandhirajan 1, Brian J. Hawkins 2, Muniswamy Madesh 1
1Department of Biochemistry, Temple University , 2Department of Anesthesiology and Pain Medicine, University of Washington

An efficient method to gain insights into visualizing the paracrine-derived ROS induction of endothelial Ca2+ signaling is described. This method takes advantage of measuring paracrine derived ROS triggered Ca2+ mobilization in vascular endothelial cells in a co-culture model.

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Biology

Multi-parameter Measurement of the Permeability Transition Pore Opening in Isolated Mouse Heart Mitochondria
Raluca Marcu 1, Chris K. Neeley 1, Georgios Karamanlidis 1, Brian J. Hawkins 1
1Department of Anesthesiology & Pain Medicine, Mitochondria and Metabolism Center, University of Washington, Seattle

A spectrofluorometric protocol for the measurement of the mitochondrial permeability transition pore opening in isolated mouse heart mitochondria is presented here. The assay involves the simultaneous measurement of mitochondria Ca2+ handling, mitochondrial membrane potential and mitochondrial volume. The procedure for obtaining high-quality and functional heart mitochondria is also described.

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