Serial frozen sections (10 microns thick) were cut parallel to the plane of the epithelium from skin, hard palate and gingiva of the pig (Sus scrofa), and beta-glucosidase activity was measured in each section. 2. In each of these tissues, there was a low constitutive level of beta-glucosidase activity in the inner portion of the epithelium, and a several-fold increase in activity was observed in the region of the stratum granulosum-stratum corneum interface. 3. The maximum specific activity of beta-glucosidase was significantly lower in gingiva (8 nmol substrate/hr/slice) than in epidermis and palate (15-18 nmol/hr/slice). 4. The increase in expression of beta-glucosidase activity near the stratum granulosum-stratum corneum boundary appears to be intimately involved in the conversion of glucosylceramides to ceramides in the final stages of differentiation. This conversion may be a major determinant of the barrier properties of the stratum corneum.