Involvement of nitric oxide in inhibitory neuromuscular transmission in equine jejunum

Peter C. Rakestraw From the Department of Surgical and Radiological Sciences, University of California, Davis, CA 95616 (Rakestraw, Snyder, Woliner), and Department of Physiology, University of Nevada, Reno, NV 89557 (Sanders, Shuttleworth).

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Jack R. Snyder From the Department of Surgical and Radiological Sciences, University of California, Davis, CA 95616 (Rakestraw, Snyder, Woliner), and Department of Physiology, University of Nevada, Reno, NV 89557 (Sanders, Shuttleworth).

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Michael J. Woliner From the Department of Surgical and Radiological Sciences, University of California, Davis, CA 95616 (Rakestraw, Snyder, Woliner), and Department of Physiology, University of Nevada, Reno, NV 89557 (Sanders, Shuttleworth).

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Kenton M. Sanders From the Department of Surgical and Radiological Sciences, University of California, Davis, CA 95616 (Rakestraw, Snyder, Woliner), and Department of Physiology, University of Nevada, Reno, NV 89557 (Sanders, Shuttleworth).

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William R. Shuttleworth From the Department of Surgical and Radiological Sciences, University of California, Davis, CA 95616 (Rakestraw, Snyder, Woliner), and Department of Physiology, University of Nevada, Reno, NV 89557 (Sanders, Shuttleworth).

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Abstract

Objectives

To evaluate the role of nitric oxide (NO), vasoactive intestinal peptide (VIP), and a transmitter acting through an apamin-sensitive mechanism in mediating inhibitory transmission in the equine jejunal circular muscle, and to determine the distribution of VIP-and NO-producing nerve fibers in the myenteric plexus and circular muscle.

Procedure

Circular muscle strips were suspended in tissue baths containing an oxygenated modified Krebs solution and attached to isometric force transducers. Responses to electrical field stimulation (EFS), tetrodotoxin, the NO antagonists l-N-nitro-arginine-methyl-ester (L-NAME) and N-nitro-l-arginine, apamin, VIP, authentic NO, and the NO donar sodium nitroprusside were tested. Immunostaining for VIP-like and NADPH diaphorase histochemical staining were performed on paraformaldehyde-fixed tissue.

Results

Subpopulations of myenteric neurons and nerve fibers in the circular muscle were positive for NADPH diaphorase and VIP-like staining. EFS caused a frequency-dependent inhibition of contractile activity. Tetrodotoxin prevented the EFS-induced inhibition of contractions. L-NAME (200 μM) and apamin 0.3 μM) significantly (P < 0.01) reduced EFS-stimulated inhibition of contractile activity at most frequencies tested. The effects of L-NAME and apamin were additive. In their combined presence, EFS induced excitation instead of inhibition (196.7% increase at 5 Hz, n = 28, P < 0.01). Inhibition of contractile activity by EFS was mimicked by sodium nitroprusside. Authentic NO (3-6 μM) abolished contractile activity. VIP induced a dose-dependent inhibition of contractile activity (89.1 ± 6.3% reduction at approximately 0.3 μM, n = 16). Antagonism of NO synthesis did not alter the response to VIP.

Conclusion

NO, VIP, and a substance acting through an apamin-sensitive mechanism appear to comediate inhibitory transmission in the equine jejunal circular muscle.

Clinical Relevance

These findings may suggest new therapeutic targets for motility disorders, such as agents that inhibit the synthesis or actions of NO. (Am J Vet Res 1996;57:1206-1213)

Abstract

Objectives

To evaluate the role of nitric oxide (NO), vasoactive intestinal peptide (VIP), and a transmitter acting through an apamin-sensitive mechanism in mediating inhibitory transmission in the equine jejunal circular muscle, and to determine the distribution of VIP-and NO-producing nerve fibers in the myenteric plexus and circular muscle.

Procedure

Circular muscle strips were suspended in tissue baths containing an oxygenated modified Krebs solution and attached to isometric force transducers. Responses to electrical field stimulation (EFS), tetrodotoxin, the NO antagonists l-N-nitro-arginine-methyl-ester (L-NAME) and N-nitro-l-arginine, apamin, VIP, authentic NO, and the NO donar sodium nitroprusside were tested. Immunostaining for VIP-like and NADPH diaphorase histochemical staining were performed on paraformaldehyde-fixed tissue.

Results

Subpopulations of myenteric neurons and nerve fibers in the circular muscle were positive for NADPH diaphorase and VIP-like staining. EFS caused a frequency-dependent inhibition of contractile activity. Tetrodotoxin prevented the EFS-induced inhibition of contractions. L-NAME (200 μM) and apamin 0.3 μM) significantly (P < 0.01) reduced EFS-stimulated inhibition of contractile activity at most frequencies tested. The effects of L-NAME and apamin were additive. In their combined presence, EFS induced excitation instead of inhibition (196.7% increase at 5 Hz, n = 28, P < 0.01). Inhibition of contractile activity by EFS was mimicked by sodium nitroprusside. Authentic NO (3-6 μM) abolished contractile activity. VIP induced a dose-dependent inhibition of contractile activity (89.1 ± 6.3% reduction at approximately 0.3 μM, n = 16). Antagonism of NO synthesis did not alter the response to VIP.

Conclusion

NO, VIP, and a substance acting through an apamin-sensitive mechanism appear to comediate inhibitory transmission in the equine jejunal circular muscle.

Clinical Relevance

These findings may suggest new therapeutic targets for motility disorders, such as agents that inhibit the synthesis or actions of NO. (Am J Vet Res 1996;57:1206-1213)

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