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Am J Physiol Cell Physiol 280: C1412-C1421, 2001;
0363-6143/01 $5.00
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Vol. 280, Issue 6, C1412-C1421, June 2001

Calcium-sensing receptor activation induces intracellular calcium oscillations

Gerda E. Breitwieser and Lucio Gama

Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205

Parathyroid hormone secretion is exquisitely sensitive to small changes in serum Ca2+ concentration, and these responses are transduced via the Ca2+-sensing receptor (CaR). We utilized heterologous expression in HEK-293 cells to determine the effects of small, physiologically relevant perturbations in extracellular Ca2+ on CaR signaling via phosphatidylinositol-phospholipase C, using changes in fura 2 fluorescence to quantify intracellular Ca2+. Chronic exposure of CaR-transfected cells to Ca2+ in the range from 0.5 to 3 mM modulated the resting intracellular Ca2+ concentration and the subsequent cellular responses to acute extracellular Ca2+ perturbations but had no effect on thapsigargin-sensitive Ca2+ stores. Modest, physiologically relevant increases in extracellular Ca2+ concentration (0.5 mM increments) caused sustained (30-40 min) low-frequency oscillations of intracellular Ca2+ (~45 s peak to peak interval). Oscillations were eliminated by 1 µM thapsigargin but were insensitive to protein kinase inhibitors (staurosporine, KN-93, or bisindolylmaleimide I). Staurosporine did increase the fraction of cells oscillating at a given extracellular Ca2+ concentration. Serum Ca2+ concentrations thus chronically regulate cells expressing CaR, and small perturbations in extracellular Ca2+ alter both resting intracellular Ca2+ as well as Ca2+ dynamics.

fura 2; intracellular calcium; calcium oscillations; thapsigargin-sensitive calcium stores; HEK-293 cells; protein kinase inhibitors


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