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Department of Pharmacology, University of Vermont, Burlington, Vermont 05405
Ca2+ sparks have been
previously described in isolated smooth muscle cells. Here we present
the first measurements of local Ca2+ transients
("Ca2+ sparks") in an intact
smooth muscle preparation. Ca2+
sparks appear to result from the opening of ryanodine-sensitive Ca2+ release (RyR) channels in the
sarcoplasmic reticulum (SR). Intracellular Ca2+ concentration
([Ca2+]i)
was measured in intact cerebral arteries (40-150 µm in diameter) from rats, using the fluorescent
Ca2+ indicator fluo 3 and a laser
scanning confocal microscope. Membrane potential depolarization by
elevation of external K+ from 6 to
30 mM increased Ca2+ spark
frequency (4.3-fold) and amplitude (~2-fold) as well as global
arterial wall
[Ca2+]i
(~1.7-fold). The half time of decay (~50 ms) was not affected by
membrane potential depolarization. Ryanodine (10 µM), which inhibits
RyR channels and Ca2+ sparks in
isolated cells, and thapsigargin (100 nM), which indirectly inhibits
RyR channels by blocking the SR
Ca2+-ATPase, completely inhibited
Ca2+ sparks in intact cerebral
arteries. Diltiazem, an inhibitor of voltage-dependent
Ca2+ channels, lowered global
[Ca2+]i
and Ca2+ spark frequency and
amplitude in intact cerebral arteries in a concentration-dependent
manner. The frequency of Ca2+
sparks (<1
s
1 · cell1),
even under conditions of steady depolarization, was too low to
contribute significant amounts of
Ca2+ to global
Ca2+ in intact arteries. These
results provide direct evidence that Ca2+ sparks exist in quiescent
smooth muscle cells in intact arteries and that changes of membrane
potential that would simulate physiological changes modulate both
Ca2+ spark frequency and amplitude
in arterial smooth muscle.
ryanodine receptor; membrane potential; calcium channels
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