TY - JOUR

T1 - Dynamics of Ca2+i and pHi in Ehrlich ascites tumor cells after Ca2+-mobilizing agonists or exposure to hypertonic solution

AU - Pedersen, Stine F.

AU - Jørgensen, Nanna K.

AU - Hoffmann, Else Kay

N1 - Key words Bradykinin - Cell volume - Na+/H+ exchanger - Simultaneous measurements - Thrombin

PY - 1998

Y1 - 1998

N2 - Intracellular free calcium concentration ([Ca2+]i) and intracellular pH (pHi) were monitored in Ehrlich ascites tumor cells using Fura-2 or 2',7',-bis-(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF), or both probes in combination. An increase in [Ca2+]i induced by thrombin or bradykinin, agonists known to elicit transient cell shrinkage in these cells, evoked a transient intracellular acidification, followed by an alkalinization. The latter was due to activation of a Na+/H+ exchanger and was inhibited under conditions preventing agonist-induced cell shrinkage without preventing the increase in [Ca2+]i. In contrast, a smaller, slower increase in [Ca2+]i elicited by thapsigargin did not cause cell shrinkage, and did not activate the Na+/H+ exchanger. Exposure to hypertonic solution was not associated with an increase in [Ca2+]i, but elicited an intracellular alkalinization similar to that induced by thrombin or bradykinin, via activation of the Na+/H+ exchanger. Thus, activation of the exchanger by the Ca2+-mobilizing agonists is suggested to be secondary to the cell shrinkage induced by these compounds. NH4Cl-induced intracellular alkalinization resulted in an increase in [Ca2+]i, apparently via stimulation of Ca2+ influx, whereas shrinkage-induced intracellular alkalinization did not stimulate Ca2+ influx. Thus, cell shrinkage appears to inhibit the Ca2+ influx otherwise resulting from alkalosis. In agreement with that notion, thapsigargin-induced Ca2+ influx was inhibited by cell shrinkage.

AB - Intracellular free calcium concentration ([Ca2+]i) and intracellular pH (pHi) were monitored in Ehrlich ascites tumor cells using Fura-2 or 2',7',-bis-(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF), or both probes in combination. An increase in [Ca2+]i induced by thrombin or bradykinin, agonists known to elicit transient cell shrinkage in these cells, evoked a transient intracellular acidification, followed by an alkalinization. The latter was due to activation of a Na+/H+ exchanger and was inhibited under conditions preventing agonist-induced cell shrinkage without preventing the increase in [Ca2+]i. In contrast, a smaller, slower increase in [Ca2+]i elicited by thapsigargin did not cause cell shrinkage, and did not activate the Na+/H+ exchanger. Exposure to hypertonic solution was not associated with an increase in [Ca2+]i, but elicited an intracellular alkalinization similar to that induced by thrombin or bradykinin, via activation of the Na+/H+ exchanger. Thus, activation of the exchanger by the Ca2+-mobilizing agonists is suggested to be secondary to the cell shrinkage induced by these compounds. NH4Cl-induced intracellular alkalinization resulted in an increase in [Ca2+]i, apparently via stimulation of Ca2+ influx, whereas shrinkage-induced intracellular alkalinization did not stimulate Ca2+ influx. Thus, cell shrinkage appears to inhibit the Ca2+ influx otherwise resulting from alkalosis. In agreement with that notion, thapsigargin-induced Ca2+ influx was inhibited by cell shrinkage.

U2 - 10.1007/s004240050623

DO - 10.1007/s004240050623

M3 - Journal article

VL - 436

SP - 199

EP - 210

JO - Pflügers Archiv - European Journal of Physiology

JF - Pflügers Archiv - European Journal of Physiology

SN - 0031-6768

IS - 2

ER -