Combined force and voltage measurement in rapidly superfused guinea pig heart cells.
We describe the construction and use of a setup that allows the rapid exchange of the solution surrounding an isolated guinea pig heart cell while simultaneously measuring the isometric force and membrane potential (Em). Cells were stably attached, by means of poly-L-lysine, to a force transducer which was adapted from one previously used for a study of frog atrial cells [N. Shepherd and F. Kavaler.Am. J. Physiol. 251 (Cell Physiol. 20): C653-C661, 1986]. The modified transducer is simple to construct and use and can be readily added to existing patch-clamp setups. The strength of attachment of a cell to the transducer exceeded the strength of the gigaseal in all of the experiments. The membrane potential was measured by means of patch electrodes and a high-impedance voltage follower. Rapidly changing extracellular K concentration [( K]o) from 5.4 to 10.8 mM caused a positive change of Em by 16.5 +/- 1.4 mV with a half-time (t1/2) of 27 +/- 4 ms. Replacing calcium in the perfusate by magnesium instantly abolished the contraction and shortened the action potential. Twitch tension returned stepwise to the control value on return of calcium to the perfusate. Our initial observations show that the patch electrode can be used successfully in conjunction with the isometric force transducer and rapid extracellular solution changes for studies of excitation and contraction coupling in isolated mammalian heart cells.
Shepherd
N
N
Research Service, Veterans Affairs Medical Center, New York, New York.
Fisher
V J
VJ
D000200:Action Potentials; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D004594:Electrophysiology / Q000295:instrumentation / Q000379:methods; D006168:Guinea Pigs; D006321:Heart / Q000502:physiology*; D006352:Heart Ventricles / Q000187:drug effects; D066298:In Vitro Techniques; D008564:Membrane Potentials / Q000187:drug effects; D009200:Myocardial Contraction / Q000187:drug effects; D010477:Perfusion / Q000295:instrumentation / Q000379:methods; D016276:Ventricular Function
D002118:Calcium