We report observations on transport of the hydrolysis-resistant dipeptide glycylsarcosine by rings of everted hamster jejunum in vitro in the presence and absence of Na+, using several substituents for Na+: Li+, K+, Cs+, Tris, choline and mannitol. At most concentrations, mediated influx of glycylsarcosine was depressed by Li+, K+, Cs+ and Tris, though not abolished. At high concentrations, it was moderately increased by choline and mannitol. Under conditions in which the tissue could concentrate the peptide in the presence of Na+, uptake was greatly depressed by all the substituents and the ability to concentrate was abolished. The Kt of mediated influx was affected in different ways according to the substituent used. Kt was reduced by Li+ replacement of Na+ and increased by choline replacement. Vmax was greatly reduced by all metallic substituents but not by non-metallic substituents. Though the results cannot yet be satisfactorily interpreted, they suggest possible reasons for previous conflicting results and show that it is impossible to make the unqualified statement that transport of glycylsarcosine is 'Na+-dependent'. It is now doubtful whether transport of this peptide into the intestinal cells is by co-transport with Na+, and the whole matter of Na+-dependence of intestinal peptide transport is in question.