Regulation of cytosolic free calcium in isolated perfused proximal tubules of Necturus

Abstract

To study the role of intracellular Ca2+ in regulating renal tubular transport of ions and water, cytosolic calcium ion activity (aiCa), cytosolic sodium ion activity (aiNa), and intracellular pH (pHi) in cells of isolated perfused proximal tubules of Necturus kidney were measured with Ca2+-, Na+-, and H+-selective microelectrodes, respectively. In control conditions, i.e., HCO3-Ringer solution on both sides of the epithelium, aiCa was 82 +/- 7 (SE) nM (n = 54), aiNa averaged 12.8 +/- 0.4 mM (n = 53), and pHi was 7.33 +/- 0.03 (n = 27). When the Na-K pump was inhibited by nominally K-free Ringer circumfusion, aiCa increased from a control level of 75 +/- 13 to 237 +/- 40 nM (paired t test; n = 16; P less than 0.001); in a different set of tubules, aiNa rose from 11.3 +/- 0.6 to 51.5 +/- 5.8 mM (n = 11; P less than 0.001). When organic solutes were deleted in the luminal perfusate, aiCa decreased from 73 +/- 11 to 61 +/- 11 nM (n = 9; P less than 0.001) and aiNa decreased from 14.6 +/- 0.6 to 8.3 +/- 0.7 mM (n = 9; P less than 0.001). Depolarization of the peritubular cell membrane with high-K, low-Na Ringer decreased aiCa from 90 +/- 12 to 55 +/- 9 nM (n = 13; P less than 0.001) and reduced aiNa from 13.1 +/- 1.0 to 7.5 +/- 0.6 mM (n = 16; P less than 0.001). Ionomycin (2 X 10(-6) M) increased aiCa from 67 +/- 10 to 158 +/- 26 nM (n = 10; P less than 0.001) and pHi from 7.33 +/- 0.03 to 7.39 +/- 0.03 (n = 27; P less than 0.001) but reduced aiNa from 11.8 +/- 0.9 to 10.3 +/- 0.7 mM (n = 11; P less than 0.001). The data are consistent with the view that aiCa is determined, in part, by the magnitude of the electrochemical potential gradient for Na ions across the basolateral cell membrane.