Glucose carriers have been shown to serve as water channels in macrophages and in oocytes injected with messenger ribonucleic acid (mRNA) encoding the glucose carrier protein (Fischbarg et al. The contribution, therefore, of glucose carriers to osmotic water permeability (Pf) in renal and intestinal epithelial cells was investigated. Pf of brush border membrane vesicles (BBMVs) and of basolateral membrane vesicles (BLMVs) was studied using stopped-flow spectrophotometry. Osmotic shrinkage of renal vesicles exhibited fast and slow components at 4 degrees C and 37 degrees C. The fast component could be inhibited by HgCl2 or dimethylsulphoxide (DMSO) at these temperatures, whereas the slow component was inhibited only at 4 degrees C. Osmotic shrinkage of intestinal BBMVs and BLMVs was homogeneous at 4 degrees C and 37 degrees C and was slightly inhibitable by HgCl2 or DMSO at 4 degrees C but not 37 degrees C. In both tissues, vesicle uptake of glucose was sensitive to HgCl2, but not to DMSO. Phlorizin and phloretin inhibited D-glucose uptake in BBMVs and BLMVs respectively, but had no significant effect on Pf. In membrane vesicles of kidney origin, Pf was tenfold higher than in membranes from intestine. This difference was not reflected by the phlorizin- and phloretin-sensitive D-glucose uptakes. Our study concludes that glucose transporters do not serve as water channels in kidney or intestine. Although membrane proteins contribute slightly to Pf at 4 degrees C, this contribution is insignificant at 37 degrees C. A membrane protein serving specifically as a water channel could only be demonstrated in renal cortical membranes.