Characteristics of calcium uptake by BeWo cells, a human trophoblast cell line

Placental transfer of maternal calcium (Ca(2+)) is a crucial process for fetal development although the biochemical mechanisms responsible for this transfer are largely unknown. We have investigated the characteristics of Ca(2+)uptake by the human placental trophoblast cell line BeWo. The kinetics studies revealed an active extracellular Ca(2+)uptake by BeWo cells, which was rapid in the first 2 min (initial velocity (V(i)) of 4.17+/-0.25 nmol/mg/min), and showed a subsequent plateau. Uptake experiments performed at V(i)with increasing concentrations of Ca(2+)resulted in a typical saturation curve (K(m)of 0.54+/-0.07 m m and V(max)of 7.07+/-0.28 nmol/mg protein/min). Lowering the pH of the incubation medium from 7.4 to 5.5 led to Ca(2+)uptake inhibition of 40-50 per cent. The presence of voltage-sensitive (l -type) Ca(2+)channels in BeWo cells was demonstrated by Western blot. Therefore, the implication of such channels in basal Ca(2+)uptake of BeWo cells was investigated. Cell depolarization with extracellular high potassium concentration (40 m m), and hyperpolarization with extracellular high chloride concentration (60 m m) or with valinomycin (10 microm) did not influence the basal Ca(2+)uptake of BeWo cells. The L-type Ca(2+)channel modulators (Bay K 8644 and Nitrendipine) had no effect on the Ca(2+)uptake. An antagonist of receptor-mediated, store-operated and voltage-gated Ca(2+)channels (SKF-96365) also did not modulate the Ca(2+)uptake of BeWo cells. Therefore, our results indicate that the basal Ca(2+)uptake of BeWo cells is inhibited by lowering pH of the incubation medium, is voltage independent, and is not influenced by l -type Ca(2+)channel and capacitative Ca(2+)conductance modulators.