Gastric parietal cell apical membranes must protect the cell from the extremely low pH and wide variations in osmolality of the gastric juice. To characterize the permeability properties of gastric apical membranes, we have measured passive permeabilities to water, protons, NH3, and small nonelectrolytes of membrane vesicles derived from parietal cells of fasted animals and fed animals. Both preparations are known to be highly enriched in H+/K(+)-ATPase, the enzyme responsible for acidifying the gastric contents. The preparations behaved as single populations, and their permeability properties were similar in all respects, permitting pooling of the results. This similarity suggests that insertion of tubulovesicles into the apical membrane does not change the behavior of the lipid bilayer. Osmotic water permeability (Pf) averaged (mean +/- SD) (2.8 +/- 0.3) x 10(-4) cm/s, a value 10-fold lower than that obtained in lecithin large unilamellar vesicles (LUV) and similar to that obtained in other water-tight epithelia. Similarly, ammonia permeability (PNH3) was low [(4.4 +/- 2.3) x 10(-3) cm/s] and 10 times below that of lecithin LUV. By contrast, proton permeability (PH+) was surprisingly high (0.030 +/- 0.011 cm/s) and similar to that of lecithin LUV. These results suggest that the pathway for proton permeation differs from that of water and NH3. Nonelectrolyte permeabilities were strikingly similar to those obtained in another water-tight epithelium, the toad urinary bladder. Moreover, these permeabilities followed Overton's rule in that permeability varied in accordance with the oil-water partition coefficient. We conclude that the gastric apical membrane, like that of several renal epithelia, is relatively water-tight and exhibits low permeabilities to small nonelectrolytes. These properties are likely to be essential to the ability of this membrane to perform its barrier function.