EMGs were recorded from the orbicularis oculi, retractor bulbi and superior rectus muscles in rabbits to investigate the time course of muscle activation during unconditioned and conditioned eye blinks. EMGs from the three muscles showed two responses, with the responses of the orbicularis oculi and retractor bulbi showing the same latency, and the responses of the superior rectus lagging. The latency of responses to periorbital electrostimulation was about 5 ms, and to air puff stimulation about 10 ms. Results showed a tight coupling of activity between muscles, with cross-correlograms peaking at 0.65 to 0.85 and showing little time shift. Stimulus-response curves showed clear non-linearities in the response of the muscle to changes in stimulus strength. Local anesthesia of the cornea had little effect on unconditionally evoked responses. The form of unconditionally evoked responses was similar with periorbital electrostimulation and air puff stimuli but differed in latency. These results show the form of the eye blink reflex response and will be of importance in interpreting electrophysiological studies of the classically conditioned eye blink of rabbits.