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Department of Pharmacology, St George's Hospital Medical School, London.

1. Sodium-calcium exchange was studied in single enzymatically isolated cells of the guinea-pig ureter using the Ca2(+)-sensitive fluorescent dye Indo-1 to monitor the intracellular Ca2+ concentration ([Ca2+]i). Patch pipettes containing Indo-1 were used to introduce the dye into cells, to set the intracellular Na+ concentration ([Na+]i) and control the membrane potential during experiments. 2. With [Na+]i set at 11-12 mM and a membrane potential of -60 or -70 mV, brief depolarization of ureter cells elicited typical voltage-gated inward currents associated with rapid increases in [Ca2+]i which showed a bell-shaped potential dependence. If Ca2+ currents were blocked with nifedipine, depolarization led to slower rises in [Ca2+]i. The rates and amplitudes of these increased monotonically with progressively larger depolarizations up to +120 mV. 3. The nifedipine-resistant rises in [Ca2+]i elicited by depolarization were potentiated when the extracellular sodium concentration ([Na+]o) was reduced. Basal levels of [Ca2+]i also increased as [Na+]o was reduced, although the dependence of this effect on [Na+]o was smaller than would be predicted if [Ca2+]i was set only by a Na(+)-Ca2+ exchange process. 4. The nifedipine-insensitive rises in [Ca2+]i elicited by depolarization were potentiated at higher basal levels of [Ca2+]i. 5. The ability of cells to reduce [Ca2+]i rapidly following Ca2+ loading during voltage-gated transients was markedly inhibited if the Na+ concentration gradient was reversed, but was little affected if the Na+ gradient was decreased by 25 or 50%. Recovery from a Ca2+ load caused by reversal of the Na+ gradient could be induced by removal of Cao2+ in the continuing absence of Nao+, indicating the importance of a Na(+)-independent [Ca2+]i-lowering system. 6. The results demonstrate that Na(+)-Ca2+ exchange can modulate [Ca2+]i when [Na+]i and the membrane potential are set at or near their physiological levels in these smooth muscle cells. [Ca2+]i does not, however, appear to be markedly sensitive to the Na+ concentration gradient under the conditions employed for these experiments, suggesting that a Na(+)-independent Ca2+ extrusion system is mainly responsible for regulating [Ca2+]i under normal conditions.

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