Rhythmic jaw muscle activities frequently occur during non-rapid eye movement sleep in patients with sleep bruxism. The present study aimed to investigate the response characteristics of the masticatory rhythm generation during non-rapid eye movement sleep in animals. Eleven guinea pigs were surgically prepared for polygraphic recordings by electromyography, electrooculography, electroencephalography and electrocardiography with the implantation of a stimulating electrode. Repetitive electrical microstimulations at three intensities were applied to the corticobulbar tract under freely moving conditions. The rhythmic electro-myographic responses of the digastric and masseter muscles were scored and analysed. Changes in cortical electro-encephalographic power and heart rate in association with these stimulations were quantified. Microstimulations to the corticobulbar tract induced rhythmic jaw muscle activities in digastric muscles, occasionally with masseter activities during wakefulness and non-rapid eye movement sleep. The response rate of rhythmic jaw muscle activities was significantly lower (p<0.01) and the response latency was significantly longer (p<0.01) during non-rapid eye movement sleep than during wakefulness. At higher stimulus intensities, the response rate increased and response latency decreased. The mean burst intervals of the digastric and masseter muscles were similar regardless of vigilance states and stimulus intensities. Induced rhythmic jaw muscle activities during non-rapid eye movement sleep were followed by a transient decrease in delta power and increases in beta power and heart rate. During non-rapid eye movement sleep, the masticatory motor system is able to generate rhythmic outputs to the jaw muscles in response to facilitatory inputs although the responsiveness was decreased from wakefulness.