© 1995 American Heart Association, Inc.
Articles |
Effects of Action Potential Duration on Excitation-Contraction Coupling in Rat Ventricular Myocytes
Action Potential Voltage-Clamp Measurements
From the Departments of Medical Physiology and Medicine, The University of Calgary, Alberta, Canada.
Correspondence to W.R. Giles, 3330 Hospital Dr NW, Calgary, Alberta, Canada T2N 4N1.
Abstract Although each of the fundamental processes involved
in excitation-contraction coupling in mammalian heart has been
identified, many quantitative details remain unclear. The initial goal
of our experiments was to measure both the transmembrane
Ca2+ current, which triggers contraction, and the
Ca2+ extrusion due to
Na+-Ca2+ exchange in a single
ventricular myocyte. An action potential waveform was used as the
command for the voltage-clamp circuit, and the membrane potential,
membrane current, [Ca2+]i, and
contraction (unloaded cell shortening) were monitored simultaneously.
Ca2+-dependent membrane current during an action
potential consists of two components: (1) Ca2+
influx through L-type Ca2+ channels
(ICa-L) during the plateau of the action potential and (2)
a slow inward tail current that develops during repolarization negative
to 
-25 mV and continues during diastole. This slow inward tail
current can be abolished completely by replacement of extracellular
Na+ with Li+, suggesting that it is due
to electrogenic Na+-Ca2+ exchange. In
agreement with this, the net charge movement corresponding to the
inward component of the Ca2+-dependent current
(ICa-L) was approximately twice that during the slow inward
tail current, a finding that is predicted by a scheme in which the
Ca2+ that enters during ICa is extruded
during diastole by a 3 Na+–1 Ca2+
electrogenic exchanger. Action potential duration is known to be a
significant inotropic variable, but the quantitative relation between
changes in Ca2+ current, action potential duration,
and developed tension has not been described in a single myocyte. We
used the action potential voltage-clamp technique on ventricular
myocytes loaded with indo 1 or rhod 2, both Ca2+
indicators, to study the relation between action potential duration,
ICa-L, and cell shortening (inotropic effect). A
rapid change from a "short" to a "long" action potential
command waveform resulted in an immediate decrease in peak
ICa-L and a marked slowing of its decline (inactivation).
Prolongation of the action potential also resulted in slowly developing
increases in the magnitude of Ca2+ transients
(145±2%) and unloaded cell shortening (4.0±0.4 to 7.6±0.4 µm).
The time-dependent nature of these effects suggests that a change in
Ca2+ content (loading) of the sarcoplasmic reticulum
is responsible. Measurement of [Ca2+]i
by use of rhod 2 showed that changes in the rate of rise of the
[Ca2+]i transient (which in rat
ventricle is due to the rate of Ca2+ release from
the sarcoplasmic reticulum) were closely correlated with changes in the
magnitude and the time course of ICa-L. These findings
demonstrate that Ca2+ release from the sarcoplasmic
reticulum can be modulated by the action potential waveform as a result
of changes in ICa-L.
Key Words: action potential duration • action potential clamp • excitation-contraction coupling • Ca2+ current • Na+-Ca2+ exchange • intracellular Ca2+
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