A subscription to JoVE is required to view this content. Sign in or start your free trial.
1Department of Surgery, Davis Heart and Lung Research Institute,The Ohio State University Wexner Medical Center, 2Department of Physiology and Cell Biology, Davis Heart and Lung Research Institute,The Ohio State University Wexner Medical Center, 3Department of Molecular Biophysics and Physiology,Rush University Medical Center, 4Department of Internal Medicine, Davis Heart and Lung Research Institute,The Ohio State University Wexner Medical Center
Chapters
- 00:05Title
- 01:29Setting Up Osmotic-stress Perfusion System and Solution Preparation
- 02:00Preparing Intact Single Flexor Digitorum Brevis (FDB) Muscle Fibers from Mouse
- 05:12Fluo-4 AM Dye Loading and Ca2+ Imaging (Sparks Measurement)
- 07:39Data Analysis
- 08:14Results: Ca2+ Sparks in Young, Aged, and Diseased Mice
- 10:28Conclusion
Described here is a method to directly measure calcium sparks, the elementary units of Ca2+ release from sarcoplasmic reticulum in intact skeletal muscle fibers. This method utilizes osmotic-stress-mediated triggering of Ca2+ release from ryanodine receptor in isolated muscle fibers. The dynamics and homeostatic capacity of intracellular Ca2+ signaling can be employed to assess muscle function in health and disease.
