The effect of the protein kinase C inhibitor, staurosporine, on intrac
ellular Ca2+ homeostasis was investigated in rat mandibular salivary a
cinar cells loaded with fura-2 Fura-2 fluorescence was measured at 510
nm while the excitation wavelength was alternated between 340 nm and
380 nm. The ratio of fluorescence intensity (F-340/380) was used as an
index of [Ca2+](i). Stimulation of acinar cells with 10 mu M carbacho
l (CCh) induced a rapid increase in F-340/380 followed by a slow decre
ase to a sustained elevated level. Addition of 1 mu M staurosporine in
the presence of CCh caused a further increase in F-340/380. In Order
to examine whether the staurosporine-induced increase in F-340/380 cou
ld be attributed either to the Ca2+ entry pathway or to the mobilizati
on of Ca2+ from intracellular Ca2+ stores, 1 mu M staurosporine was ad
ded in the presence of CCh after Ca2+ had been omitted from the perfus
ate. Even in the absence of extracellular Ca2+, F-340/380 Still increa
sed slowly from 0.75 +/- 0.05 to 1.57 +/- 0.24 after a delay ranging b
etween 5 min and 10 min. However, the IP3-sensitive intracellular Ca2 stores did not seem to play a major role in this phenomenon because s
taurosporine still increased F-340/380 by 0.6 +/- 0.10 after the compl
ete depletion of IP3-sensitive Ca2+ stores by the exposure of cells to
I mu M thapsigargin, a microsomal Ca2+ ATPase inhibitor, in Ca2+-free
conditions. These results suggest that staurosporine mobilizes Ca2+ f
rom IP3-insensitive intracellular Ca2+ stores in rat mandibular saliva
ry acinar cells.