The superficial buffer barrier in venous smooth muscle: sarcoplasmic reticulum refilling and unloading.
1. The interaction of Ca2+ transport in the plasmalemma and the sarcoplasmic reticulum (SR) was investigated in smooth muscle of the rabbit inferior vena cava. We tested the possibility of direct refilling of the SR with extracellular Ca2+ and of the existence of a vectorial Ca2+ extrusion pathway from the SR lumen to the extracellular space suggested by earlier results. 2. After depletion with caffeine the SR was loaded with Ca2+ to increasing levels by incubation in a high potassium 1.5 mM Ca2+ solution and a 10 mM Ca2+ zero Na+ solution, respectively. Thapsigargin, 2 microM, (a specific SR Ca(2+)-ATPase blocker) completely blocked refilling of the SR in either of the above solutions, indicating that the SR Ca(2+)-ATPase is essential for this process. 3. Three different agents, caffeine, ryanodine and thapsigargin, which inhibit Ca2+ accumulation by the SR, increased the steady state intracellular Ca2+ concentration in the rabbit inferior vena cava. 4. Measurements of Mn2+ induced quenching of the intracellular fura-2 signal during pharmacological manipulation of the SR content showed that these three agents did not stimulate divalent cation entry. 5. On the other hand, stimulation with noradrenaline caused a marked increase in Mn2+ influx, which was blocked by 2 mM Ni2+. Mn2+ entry stimulated by high K+ solution was blocked by 1 microM diltiazem. 6. We conclude that the SR refilling has to be mediated by the SR Ca(2+)-ATPase. Inhibition of Ca2+ accumulation by the SR causes an increase in the steady state intracellular Ca2+ concentration. This observation cannot be explained by an increase in Ca2+ influx into the smooth muscle cells of the rabbit inferior vena cava. Alternatively these results suggest the existence of a continuous vectorial release of Ca2+ from the SR lumen to the extracellular space.
Chen
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University of Miami School of Medicine, Department of Molecular and Cellular Pharmacology, FL 33101.
van Breemen
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C
D000818:Animals; D002021:Buffers; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D002412:Cations / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D066298:In Vitro Techniques; D008297:Male; D008345:Manganese / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D009638:Norepinephrine / Q000494:pharmacology; D011817:Rabbits; D012433:Ryanodine / Q000494:pharmacology; D012519:Sarcoplasmic Reticulum / Q000201:enzymology / Q000378:metabolism*; D013729:Terpenes / Q000494:pharmacology; D019284:Thapsigargin; D014682:Vena Cava, Inferior / Q000187:drug effects
D002021:Buffers; D002412:Cations; D013729:Terpenes; D012433:Ryanodine; D002110:Caffeine; D008345:Manganese; D019284:Thapsigargin; D000252:Calcium-Transporting ATPases; D002118:Calcium; D009638:Norepinephrine