U. Tiemann et al., EFFECT OF ATP AND PLATELET-ACTIVATING-FACTOR ON INTRACELLULAR CALCIUMCONCENTRATIONS OF CULTURED OVIDUCTAL CELLS FROM COWS, Journal of Reproduction and Fertility, 108(1), 1996, pp. 1-9
Oviductal endosalpingeal cells were mechanically isolated from heifers
at different reproductive stages (cyclic: days 8-14, at oestrus: day
0 and pregnant: day 7) and cultured until 100% confluent. The cells we
re loaded with the Ca2+-sensitive fluorochrome fura-2/AM, and cytosoli
c free calcium ([Ca2+](i)) was monitored spectrofluorimetrically and b
y use of a microscope image analysis system. ATP (400 mu mol l(-1)) ev
oked a prompt increase in [Ca2+](i) in all cell preparations in both t
he presence or absence of extracellular Ca2+ when measured with the cu
vette method. Single cell measurements using oviductal cells from cycl
ic heifers revealed a heterogeneous [Ca2+](i) pattern in response to A
TP, with some cells either failing to respond or reacting very slowly.
Platelet-activating factor (PAF, 30 nmol l(-1)) induced a rapid incre
ase in [Ca2+](i) especially in cells derived from cyclic and pregnant
animals, but the effect was significantly less in cells from heifers a
t oestrus. The increase in [Ca2+](i) in bovine cells induced by PAF wa
s reduced when extracellular calcium was depleted, indicating that the
effect of PAF on [Ca2+](i) involves an influx from the extracellular
space. Voltage-sensitive calcium channels do not appear to be involved
in the influx of extracellular Ca2+ since verapamil had no effect on
the PAF-induced increase in [Ca2+](i). The PAF receptor antagonist WEB
2086 inhibited the PAF-mediated effects on [Ca2+](i).