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Vollum Institute, Oregon Health Sciences University, Portland 97201.

1. The role of the endothelium as an effector of the neurogenic cholinergic vasodilatation in submucosal arterioles of the guinea-pig ileum was investigated by measuring changes in arteriolar diameter in response to exogenous application of muscarine or electrical stimulation of the submucosal ganglia. 2. NG-Monomethyl-L-arginine (L-NMMA), an inhibitor of nitric oxide (NO) synthesis, competitively inhibited the vasodilatation produced by muscarine in arterioles which had been preconstricted with the prostaglandin analogue U46619. L-Arginine (10 mM), but not D-arginine (10 mM), prevented the inhibition by L-NMMA. 3. Neither tetrodotoxin (TTX, 1 microM), nor the cyclo-oxygenase inhibitor, indomethacin (10 microM), altered the muscarinic vasodilatation or the inhibitory effect of L-NMMA. 4. Sodium nitroprusside (SNP), an activator of the soluble guanylate cyclase, dilated the arterioles in a concentration-dependent manner. This vasodilatation was unaffected by L-NMMA but was abolished by the guanylate cyclase inhibitor, methylene blue (10 microM). In addition, methylene blue antagonized the muscarinic vasodilatation to a similar degree as did L-NMMA. 5. The vasodilatation produced by ganglionic stimulation (10 Hz, 10 s) was blocked by TTX and the muscarinic receptor antagonist, 4-diphenylacetoxy-N-methyl-piperidine methiodide (4-DAMP, 1 microM). The neurally evoked vasodilatation was inhibited by 70% in the presence of L-NMMA; this inhibition was prevented by L-arginine. Methylene blue inhibited the neurogenic vasodilatation to the same extent as did L-NMMA. 6. These results show that arteriolar vasodilatation by muscarine is mediated mainly through the release of NO formed from L-arginine; the origin of the L-arginine appears to be the endothelium. These results also demonstrate that acetylcholine released from submucosal nerves onto submucosal blood vessels reaches the endothelium to cause the release of NO formed from L-arginine; the endothelial-derived NO dilates the arteriole.

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