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1. The role of the sarcoplasmic reticulum (SR) was investigated in spontaneous and agonist-induced uterine Ca2+ transients, by combining low- (mag-fluo-4) and high-affinity (fura-2) indicators to measure intraluminal SR ([Ca2+]L) and cytosolic ([Ca2+]i) calcium concentration, simultaneously, in single smooth muscle cells from pregnant rat uterus.
2. Carbachol or ATP, in the absence of extracellular Ca2+, decreased [Ca2+]L and increased [Ca2+]i. Although some replenishment (around 50 %) occurred in its absence, extracellular Ca2+ was required for full replenishment of the SR Ca2+.
3. In 4/15 cells, ATP evoked oscillations of [Ca2+]i. These were accompanied by successive release and re-uptake of SR Ca2+. Inhibition of the SR Ca2+-ATPase with thapsigargin abolished the oscillations and luminal changes.
4. Spontaneous [Ca2+]i transients produced no detectable changes in [Ca2+]L. The larger [Ca2+]i transients evoked by high-K+ depolarisation increased [Ca2+]L. Spontaneous activity was inhibited when [Ca2+]L was increased.
5. These data show that it is possible to simultaneously measure SR and cytosolic [Ca2+], and to investigate their response to agonist application and spontaneous activity.
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