The C6 rat glioma cell line is broadly used as a model in studies of glial cell differentiation. In the present study we demonstrated a significantly higher total cellular, but especially membrane-associated, activity of dipeptidyl peptidase IV in differentiated C6 cells in comparison with their proliferating counterparts. The majority, but not all, of enzyme isoelectric focusing isoforms from differentiated C6 cells displayed a substantially higher activity compared to the proliferating cells, with G-P-NHMec as the substrate. Non-denaturing polyacrylamide gradient gel electrophoresis showed the presence of one major peak of activity, dipeptidyl peptidase IV (Mr of about 220000), in both proliferating and differentiated C6 cells. The results indicate that dipeptidyl peptidase IV regulation is associated with C6 rat glioma cell differentiation.