Chloride conductance in membrane vesicles from human placenta using a fluorescent probe. Implications for cystic fibrosis

Abstract

Previous evidence suggests that the molecular defect in cystic fibrosis (CF) could reside in an altered chloride conductance of epithelial tissues. Since the brush border of the syncytiotrophoblast of the chorionic villi of human placenta is on abundant source of epithelial membranes and it is unaltered by secondary pathology or treatment we chose to characterize its chloride conductance and to compare it in normal and CF membranes. Chloride transport was studied in microvillar vesicles (MVV) by the quenching of the fluorescent probe 6-methoxy- N-(3-sulfopropyl)quinolinium (SPQ).Chloride conductance at 23°C: (a) increased by 39% under a membrane potential change of 70 mV; (b) was inhibited by diphenylamine 2-car☐ylate ( K l × 150 μM); (c) displayed an activation energy of 3.5 kcal · mol −1. The comparison of the chloride conductance for an inwardly directed gradient of 150 mM Cl − at 23° C (membrane potential set at 0 mV) between CF and control membranes was not significantly different. These findings demonstrate the presence of a chloride conductive pathway in microvillar vesicles from human placenta and preliminary results exclude major differences in the conductance of CF derived material in the absence of neurohormonal stimuli.