Acupuncture or acupuncture-like stimulation applied to different body areas can modify autonomic nerve activity to various organs, including gut, bladder, adrenal medulla, and the heart. We studied the reflex bradycardia in response to insertion into the skin and underlying muscles and twisting of an acupuncture needle in pentobarbital-anesthetized rats. We found that acupuncture-like stimulation of forelimb, hindlimb, chest, and abdomen all produced significant heart rate decreases. Rate minima were reached at the end of the 60-second stimulation episode and significant bradycardia persisted for about 40 s after stimulation ended. Heart rate decreases were paralleled by decreases in cardiac sympathetic nerve activity, and could be produced by electrical stimulation of group IV muscle afferent fibers (tibial nerve). Electrical stimulation of the tibial nerve at rates as low as 0.1-2 Hz was effective for eliciting heart rate decreases. Nerve fiber groups were defined by stimulation of and recording from tibial nerve. Activation of groups I, II, or III fiber was ineffective for eliciting the reflex bradycardia. Sympathectomy, high spinal transection, or infusion of the GABA(A) receptor antagonist, bicuculline, into the cisterna magna were all effective for disrupting the reflex bradycardia. Vagotomy and opioid receptor blockade were ineffective for disrupting the reflex pathway. We conclude that the reflex pathway to decrease heart rate by acupuncture-like stimulation consists of mainly group IV muscle afferent fibers whose activity (even very low rate of activity) leads to the activation of GABA-ergic neurons in the brainstem and an inhibition of sympathetic outflow to the heart.