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Volume 270, Number 28, Issue of July 14, pp. 16666-16670, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Calcitonin Increases Cytosolic Free Calcium Concentration via Capacitative Calcium Influx

Anna Teti , Rossella Paniccia , Steven R. Goldring

The calcitonin receptor has been proposed to function as an extracellular Ca concentration ([Ca]) sensor (Stroop, S. D., Thompson, D. L., Kuestner, R. E., and Moore, E. E.(1993) J. Biol. Chem. 268, 19927-19930). To test this hypothesis we studied the LLC-PK renal tubular cells and the PC cells, a cell line stably transfected with the cloned porcine calcitonin receptor. [Ca] was measured by fura-2 single cell microfluorometry. Addition to the cells equilibrated in 1.25 mM Ca-containing media of 1-10 mM extracellular Ca did not result in a significant increase of [Ca]. Treatment with 10M salmon calcitonin (sCT) elicited a rapid, persistent elevation of [Ca]. Addition of 1-10 mM extracellular Ca in the presence of sCT induced a significant [Ca]elevation, about 10-fold that observed in the absence of the hormone. Ca influx was inhibited by lanthanum. The rise of [Ca] at elevated [Ca]was not due to a Ca sensing mechanism with release of Ca from intracellular stores, since it was prolonged, and was not abolished by prior depletion of Ca stores with 10M thapsigargin. On the contrary, this agent potentiated Ca influx after addition of 1-10 mM Ca by 13-fold versus control. Prior stimulation of [Ca] with 10M arginine-vasopressin had similar effects, enhancing the subsequent Ca influx. Enhancement of Ca influx by sCT was confirmed by increased Mn quenching of fura-2 fluorescence. In conclusion, arginine-vasopressin or calcitonin enhance Ca influx in LLC-PK cells via a Ca release-activated conductance, probably dependent upon capacitative Ca entry. Thus, these effects are not unique to the calcitonin receptor and argue against the receptor functioning as a [Ca] sensor.



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