Abstract In a previous study, we found that electrical and chemical stimulation of the red nucleus (RN) suppressed the high-threshold afferent-evoked jaw-opening reflex (JOR). It has been reported that the RN receives bilaterally projection fibers from the raphe magnus nucleus (RMg), and that stimulation of the RMg inhibits the tooth pulp-evoked nociceptive JOR. These facts imply that RMg-induced inhibition of the JOR could be mediated via the RN. The present study first examines whether stimulation of the RMg suppresses the high-threshold afferent-evoked JOR. The JOR was evoked by electrical stimulation of the inferior alveolar nerve (IAN), and was recorded as the electromyographic response of the anterior belly of the digastric muscle. The stimulus intensity was 4.0 (high-threshold) times the threshold. Conditioning electrical stimulation of the RMg significantly suppressed the JOR. A further study then examined whether electrically induced lesions of the RN or microinjection of muscimol into the RN affects RMg-induced suppression of the JOR. Electrically induced lesions of the bilateral RN and microinjection of muscimol into the bilateral RN both reduced the RMg-induced suppression of the JOR. These results suggest that RMg-induced suppression of the high-threshold afferent-evoked JOR is mediated by a relay in the RN.