Inhibitory pathways in the circular muscle of rat jejunum

Conflicting data have been reported on the contribution of nitric oxide (NO) to inhibitory neurotransmission in rat jejunum. Therefore, the mechanism of relaxation and contribution to inhibitory neurotransmission of NO, adenosine 5′‐triphosphate (ATP), vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase‐activating peptide (PACAP) was examined in the circular muscle of Wistar–Han rat jejunum. Mucosa‐free circular muscle strips were precontracted with methacholine in the presence of guanethidine and exposed to electrical field stimulation (EFS) and exogenous NO, ATP, VIP and PACAP. All stimuli induced reduction of tone and inhibition of phasic motility. Only electrically induced responses were sensitive to tetrodotoxin (3 × 10−6M). NO (10−6–10−4M)‐induced concentration‐dependent relaxations that were inhibited by the soluble guanylyl cyclase inhibitor 1H‐[1,2,4]‐oxadiazolo‐[4,3‐a]‐quinoxalin‐1‐one (ODQ; 10−5M) and the small conductance Ca2+‐activated K+‐channel blocker apamin (APA; 3 × 10−8M). Relaxations elicited by exogenous ATP (10−4–10−3M) were inhibited by the P2Y purinoceptor antagonist reactive blue 2 (RB2; 3 × 10−4M), but not by APA and ODQ. The inhibitory responses evoked by 10−7M VIP and 3 × 10−8M PACAP were decreased by the selective PAC1 receptor antagonist PACAP6–38 (3 × 10−6M) and APA. The VPAC2 receptor antagonist PG99‐465 (3 × 10−7M) reduced relaxations caused by VIP, but not those by PACAP, while the VPAC1 receptor antagonist PG97‐269 (3 × 10−7M) had no influence. EFS‐induced relaxations were inhibited by the NO‐synthase inhibitor Nω‐nitro‐L‐arginine methyl ester (3 × 10−4M), ODQ and APA, but not by RB2, PG97‐269, PG99‐465 and PACAP6–38. These results suggest that NO is the main inhibitory neurotransmitter in the circular muscle of Wistar–Han rat jejunum acting through a rise in cyclic guanosine monophosphate levels and activation of small conductance Ca2+‐dependent K+ channels. British Journal of Pharmacology (2004) 143, 107–118. doi:10.1038/sj.bjp.0705918