The present study was designed to examine 1) whether there are vasodilator fibres in the masseter muscle, and 2) if there are, to establish the neural pathways mediating these responses in urethane-anaesthetized rats. Electrical stimulation of the central cut end of the lingual nerve (LN) elicited intensity- and frequency-dependent increases of the blood flow in the masseter muscle (MBF) and lower lip (LBF). Increases in both the MBF and LBF evoked by the LN stimulation were reduced by hexamethonium in a dose-dependent manner (1-10 mg kg^-1). Pretreatment with phentolamine or propranolol at a dose of 100 µg kg^-1 had no effect on the increases in either MBF or LBF evoked by LN stimulation. Pretreatment with atropine (100 µg kg^-1) significantly reduced the MBF increase induced by LN stimulation, but not that in the LBF. The sectioning the superior cervical sympathetic trunk did not affect the responses. MBF increases occurred with electrical stimulation of the trigeminal ganglion, and these increases were significantly reduced by both the administration of hexamethonium and atropine. Lidocaine microinjection into the trigeminal spinal nucleus or salivatory nuclei caused a significant attenuation of the LN-induced MBF increases. When WGA-HRP was injected into the masseter muscle, labelled neurones were abundantly observed in the otic ganglion. The present study indicates that there are parasympathetic cholinergic and non-cholinergic vasodilator fibres originating from cell bodies in otic ganglion in the rat masseter muscle. The MBF increase evoked by activation of the parasympathetic fibres occurred via the trigeminal mediated reflex, suggesting that the novel parasympathetic vasodilator response may play an important role in the regulation of the hemodynamics of jaw muscles.