Partial nicotinic receptor blockade unmasks a modulatory role of nitric oxide on urethral striated neuromuscular transmission.
The objective of this study was to investigate the possible modulatory role of endogenous nitric oxide (NO) production on the urethral striated muscle (USM) function in the sheep urethra. Significant NO synthase (NOS) activity was measured in both the particulate and cytosolic fractions of USM homogenates. NOS activity was calcium-dependent and showed greater inhibition by NOS inhibitors selective of the neural NOS isoform (nNOS). nNOS immunoreactivity was present in intramural nerves as well as in the sarcolemma of some striated fibers, being denser at the neuromuscular junction (NMJ). Double immunolabeling showed co-localization of nNOS with both alpha-bungarotoxin and choline acetyltransferase, at the USM endplates. For the first time, functional data support a role of NO on the USM contractility "in vitro," which became evident following partial nicotinic receptor inactivation with low concentrations of D-tubocurarine. Only under D-tubocurarine (0.25 microM) treatment, different NOS inhibitors, specially N(G)-propyl-L-arginine, as well as the guanylate cyclase inhibitor ODQ, all showed a significant enhancing effect on contractions induced by electrical field stimulation of intrinsic somatic nerves. These data suggest that local production of NO at the urethral NMJ may modulate release and/or action of acetylcholine on motor endplates by cyclic GMP-mediated effects. This modulatory action could be especially relevant when neuromuscular transmission at the USM is impaired.
Department of Physiology, Veterinary School, Complutense University, Madrid, Spain.
A
A
García-Pascual
G
G
Costa
A
A
Labadía
E
E
Jimenez
D
D
Triguero
E
E
Rodríguez-Veiga
J
J
González-Soriano
D000818:Animals; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D004558:Electric Stimulation; D005260:Female; D006162:Guanylate Cyclase / Q000502:physiology; D009045:Motor Endplate / Q000201:enzymology; D018482:Muscle, Skeletal / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000032:analysis*; D009469:Neuromuscular Junction; D009569:Nitric Oxide / Q000096:biosynthesis* / Q000502:physiology; D019001:Nitric Oxide Synthase / Q000032:analysis*; D052248:Nitric Oxide Synthase Type I; D011978:Receptors, Nicotinic / Q000187:drug effects*; D012756:Sheep; D014018:Tissue Distribution; D014403:Tubocurarine / Q000494:pharmacology; D014521:Urethra / Q000502:physiology*; D014553:Urinary Tract Physiological Phenomena
D009419:Nerve Tissue Proteins; D011978:Receptors, Nicotinic; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I; D002795:Choline O-Acetyltransferase; D006162:Guanylate Cyclase; D014403:Tubocurarine