We examined the enhancing action of sodium caprate and palmitoylcarnitine on the permeability of fluorescein isothiocyanate dextran 4000 as a paracellular permeant compound in isolated rat and human colon samples using the Ussing-type chamber method. In the absence of an enhancer, the permeation clearance of fluorescein isothiocyanate dextran 4000 was not significantly different in the rat and human colons, but the electric membrane resistance was smaller in the rat colon than in the human colon. Sodium caprate and palmitoylcarnitine increased permeation clearance and decreased electric membrane resistance in both types of colonic membrane, showing that the rat colon can be used as a model of the human colon for studies of enhancer effects. A calmodulin antagonist significantly inhibited the action of sodium caprate in both colonic membranes. However, it tended to promote the effects of palmitoylcarnitine on permeation clearance and electric membrane resistance. These results suggest that sodium caprate induces the contraction of the perijunctional actomyosin ring to widen the tight junction and that the mechanism of palmitoylcarnitine is different from that of sodium caprate in the human colon, as reported previously for Caco-2 cell monolayers.