University of Washington, Seattle

2 ARTICLES PUBLISHED IN JoVE

Biology

Visualization of Vascular Ca²⁺ Signaling Triggered by Paracrine Derived ROS

Karthik Mallilankaraman¹, Rajesh Kumar Gandhirajan¹, Brian J. Hawkins², Muniswamy Madesh¹

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.

Biology

Multi-parameter Measurement of the Permeability Transition Pore Opening in Isolated Mouse Heart Mitochondria

Raluca Marcu¹, Chris K. Neeley¹, Georgios Karamanlidis¹, Brian J. Hawkins¹

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.