Zusammenfassung
The local calcium release flux signals (calcium spikes) evoked by
membrane depolarization were recorded at high temporal resolution
(2000 lines s(-1)) in isolated ventricular myocytes of male rats,
using combination of scanning confocal microscopy and the patch-clamp
technique. The kinetic properties of calcium spikes were investigated.
The time course of calcium spike activation could be described reliably
by a model with higher-order (n = 3) kinetics, but not by a first-order
exponential process. A model of calcium spike with calcium release
termination coupled to its activation was preferential to a model
with the release termination independent of its activation. Three
fluorescent calcium dyes (OG-5N, fluo-3, and fluo-4) were compared
for calcium spike measurements. Experimental measurements as well
as simulations showed that the occurrence and latency of calcium
spikes could be measured faithfully with all indicators, while the
kinetics of calcium spikes was reliably traced only with OG-5N. Calcium
spikes evoked by a step depolarization from -50 to 0 mV commenced
with a mean latency of 4.1 +/- 0.2 ms and peaked 6.7 +/- 0.2 ms later.
Their full amplitudes were normally distributed. The activation time
constant of calcium spikes was 3.1 +/- 0.1 ms, and the time constant
of termination was 5.5 +/- 0.2 ms. A negative correlation was observed
between the observed amplitude of calcium spikes and their time constant
of activation, but there was no correlation between their observed
amplitude and time constant of termination, in agreement with the
concept of steep calcium-dependent activation and fateful inactivation
of calcium release flux.
Nutzer