Chloride transport mechanisms in isolated plasma membrane vesicles were studied to characterize pathways for transcellular transport of chloride. Microvillous membrane (MVM) and basal membranes (BM) vesicles were isolated from term placentae. Western blot analysis of the anion exchanger isoform 1 (AE1) demonstrated that the density of AE1 was 12-fold higher on the MVM compared to the BM. At 30 sec, the Cl- uptake in the absence of a potential difference (p.d.) was 457.3 +/- 69.7 and 111.0 +/- 29.1 pmol/mg protein in MVM and BM, respectively (mean +/- SEM, n=6). Chloride transport pathways were characterized using diisothiocyano-2'2-disulphonic stilbene. (DIDS, 0.1 mM) and diphenylamine-2-carboxylate (DPC, 0.5 mM) in the absence or presence of inside positive membrane potentials. Anion exchange (DIDS-sensitive uptake at zero mV) was found in the MVM only. Both MVM and BM showed increased chloride uptake in the presence of inside positive potentials, suggesting the presence of chloride conductance pathways. The chloride uptake with a 25-mV inside positive p.d. could be inhibited by both DIDS and DPC in MVM and BM. However greater potentials (50 mV) showed no significant inhibition by DIDS or DPC in BM. In conclusion, the anion exchanger is unlikely to contribute significantly to chloride fluxes across BM. The data also suggest the presence of Cl- conductance pathways in both the MVM and BM which are sensitive to both DIDS and DPC.