Measuring Cardiac Myocyte Ca2+ and Contractility

Adult mouse ventricular myocytes were isolated following a previously described protocol (Liao and Jain, 2007 (link)). Intracellular Ca²⁺ and contractility were measured at 37°C in isolated myocytes loaded with Fura 2-AM in the absence or presence of isoproterenol (ISO) (1 μmol/L) using an IonOptix system (Milton, MA), as previously described (Guillory et al., 2013 (link)). Myocytes were paced at 1 Hz with IonOptix MyoPacer Field stimulator (pulse duration 1 ms; 5 volts). Data were analyzed using IonWizard (IonOptix) software.

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McConnell B.K., Singh S., Fan Q., Hernandez A., Portillo J.P., Reiser P.J, & Tikunova S.B. (2015). Knock-in mice harboring a Ca2+ desensitizing mutation in cardiac troponin C develop early onset dilated cardiomyopathy. Frontiers in Physiology, 6, 242.

Publication 2015

MyoPacer Field stimulator IonWizard

Adult Contractility Fura 2 am Intracellular Isoproterenol Mouse Myocytes Pulse Ventricular

Corresponding Organization :

Other organizations : University of Houston, The Ohio State University

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Variable analysis

independent variables

Isoproterenol (ISO) (1 μmol/L)

dependent variables

Intracellular Ca2+ concentration
Contractility

control variables

Temperature (37°C)
Pacing (1 Hz with 1 ms pulse duration, 5 volts)

controls

Absence of isoproterenol (ISO) (negative control)

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