Nk. Jorgensen et al., ON THE ROLE OF CALCIUM IN THE REGULATORY VOLUME DECREASE (RVD) RESPONSE IN EHRLICH MOUSE ASCITES TUMOR-CELLS, The Journal of membrane biology, 157(3), 1997, pp. 281-299
The putative role for Ca2+ entry and Ca2+ mobilization in the activati
on of the regulatory volume decrease (RVD) response has been assessed
in Ehrlich cells. Following hypotonic exposure (50% osmolarity) there
is: (i) no increase in cellular Ins(1,4,5)P-3 content, as measured in
extracts from [2-H-3] myoinositol-labeled cells, a finding at variance
with earlier reports from our group; (ii) no evidence of Ca2+-signali
ng recorded in a suspension of fura-2-loaded cells; (iii) Ca2+-signali
ng in only about 6% of the single, fura-2-loaded cells at 1-mM Ca2+ (1
% only at 0.1-mM Ca2+ and in Ca2+-free medium), as monitored by fluore
scence-ratio imaging; (iv) no effect of removing external Ca2+ upon th
e volume-induced K+ loss; (v) no significant inhibition of the RVD res
ponse in cells loaded with the Ca2+ chelator BAPTA when the BAPTA-load
ing is performed in K+ equilibrium medium; (vi) an inhibition of the s
welling-induced K+ loss (about 50%) at 1-mM Ba2+, but almost no effect
of charybdotoxin (100 nM) or of clotrimazole (10 mu M), reported inhi
bitors of the K+ loss induced by Ca2+-mobilizing agonists. Thus, Ca2signaling by Ca2+ release or Ca2+ entry appears to play no role in the
activation mechanism for the RVD response in Ehrlich cells.