J. Biol. Chem., Vol. 261, Issue 17, 7669-7674, Jun, 1986
Unusual sensitivity of cytosolic free Ca2+ to changes in extracellular Ca2+ in rat C-cells
RM Fried and AH Tashjian Jr
An essential function of C-cells is to monitor extracellular Ca2+
concentration ([Ca2+]e) and to respond to changes in [Ca2+]e by regulating
hormone secretion. Using the calcitonin-secreting rat C-cell line rMTC
44-2, we have investigated a possible tight linkage between [Ca2+]e and
cytosolic free Ca2+ ([Ca/+]i). We have demonstrated, using the Ca2+
indicator Quin 2, that the [Ca2+]i is particularly sensitive to changes in
[Ca2+]e. Sequential increases in [Ca2+]e as small as 0.1 mM evoke clear
elevations in [Ca2+]i. In contrast, other cell types tested did not alter
their [Ca2+]i in response to increasing [Ca2+]e even to levels as high as
4.0 mM. Sequential 1.0 mM increments in [Ca2+]e caused the [Ca2+]i to rise
from a base line of 357 +/- 20 nM Ca2+i at 1.0 mM Ca2+e to a maximum of
1066 +/- 149 nM Ca2+i at 5.0 mM Ca2+e. [Ca2+]e above 2.0 mM produced a
biphasic response in [Ca2+]i consisting of an immediate (less than 5 s)
spike followed by a decay to a new plateau. Treatment of rMTC 44-2 cells
with either 50 mM K+ or 100 nM ionomycin at 1.0 mM Ca2+e caused an
immediate spike in [Ca2+]i to micromolar levels. Pretreatment with EGTA or
verapamil inhibited completely the increase in [Ca2+]i induced by 50 mM K+.
However, pretreatment with EGTA only slightly attenuated the spike phase in
[Ca2+]i produced by ionomycin, demonstrating that ionomycin released
intracellular stores of calcium. We conclude that rMTC 44-2 cells regulate
[Ca2+]i by monitoring small physiological changes in [Ca2+]e, the primary
secretagogue for C-cells.