An outwardly rectifying chloride channel in BeWo choriocarcinoma cell line

Abstract

In this study, an outwardly rectifying chloride channel was characterized in the trophoblastic cell line BeWo, a human hormone-synthesizing cell which displays many biochemical and morphological properties similar to those reported for the human cytotrophoblast. Ion channel activity was recorded in the cell attached and inside-out configurations with standard patch-clamp technology. In most of the BeWo cells studied, the channel under symmetrical N-methyl-d-glucamine (NMDG-Cl) concentration (Na+ free solution) in both sides of the membrane exhibited spontaneous activity, an outwardly rectifying current/voltage relationship and single-channel conductances of 15 pS and 48 pS for inwards and outwards currents, respectively. The channel has a low permeability for gluconate with a relative permeability Pgluconate/PCl of 0.23, and a higher permeability to I−. The open probability (Po) of the channel exhibited dependence with the applied membrane potential with greater activity at positive pulses. The channel activity was inhibited by the sulphonylurea hypoglycemic agent glibenclamide (50 μM) or by diphenylamine-2-carboxylate (DPC, 500 μM) added to the cytoplasmic side of the patch whereas conductances remained unchanged. The blockade with glibenclamide and DPC was independent of the applied membrane potential. All these results are characteristic of the outwardly rectifying Cl channel (ORCC) found in other types of cells. Neither Po, conductances nor reversal potential (Er) values were affected by the absence of intracellular Ca2+, suggesting that the channel is not sensitive to Ca2+.