Previous studies have shown that serine protease inhibitors promote neurite outgrowth from neuroblastoma cells, sympathetic neurons and sensory ganglia in culture. In the present study, a neurite promoting activity of thrombin inhibitors such as hirudin, D-Phe,Pro,Arg-CH2Cl, and paraamidinophenylalanine derivatives, was found in rat embryo (E17) septal neurons in primary culture. In contrast, no effect was shown on choline acetyl-transferase activity of septal fragments in culture. These results suggest that thrombin inhibitors might interact with a thrombin-like protease involved in the control of neurite outgrowth.