This study was performed to elucidate the type of afferents that mediate the multiple actions of 5‐hydroxytryptamine (5‐HT) on mesenteric nerve discharge. Electrophysiological recordings were made from mesenteric afferents innervating the mid‐jejunum of the urethane‐anaesthetized rat. The discharge of single nerves within the whole nerve recording was monitored using waveform discrimination software.

Afferents responded to 5‐HT in one of two ways: a short latency, transient excitation mediated by 5‐HT₃ receptors, or a delayed onset, more prolonged effect that was 5‐HT_2A receptor mediated. Afferents showing the 5‐HT₃‐mediated response did not respond to luminal distension but were sensitive to intraluminal hydrochloric acid (150 mM) in twenty‐eight of twenty‐nine experiments. In eight experiments, the 5‐HT₃‐mediated response was reversibly abolished by a 2 min exposure to intraluminal application of local anaesthetic (2 % Xylocaine).

Mechanosensitive afferents which responded to distension (< 10 cmH₂O) did not show a 5‐HT₃‐mediated response (P= 0.92, n= 14), and maintained this mechanosensitivity after luminal anaesthesia. Mechanosensitive afferents did show a secondary response to 5‐HT that was significantly attenuated by atropine (100‐200 μg kg⁻¹), whereas hexamethonium (8 mg kg⁻¹) had no effect.

In animals whose vagal afferent contribution to their mesenteric nerves had been eliminated by chronic truncal vagotomy, the 5‐HT₃‐mediated response was absent in thirty‐six of thirty‐six nerve bundles. In contrast, mechanosensitivity to distension and the secondary response to 5‐HT could still be evoked.

These results suggest that 5‐HT stimulates mesenteric afferents by a direct action on 5‐HT₃ receptors that are present on vagal mucosal afferent terminals. The mucosal afferent response to luminal acid, however, was unaffected by treatment with granisetron (0.5 mg kg⁻¹) indicating that endogenous 5‐HT from enterochromaffin cells is not essential for transduction of this luminal signal. In contrast, mechanosensitivity in non‐vagal afferents was modulated by 5‐HT following an intestinal motor response which was influenced by cholinergic tone.