title: Inhibition of relaxations to nitrergic stimulation of the mouse anococcygeus by duroquinone.
abstract: 1. The role of copper/zinc superoxide dismutase (Cu/Zn SOD) in protection of nitrergic neurotransmission in the mouse anococcygeus was investigated by use of duroquinone (DQ), which generates superoxide anions within tissues via reduction by flavoprotein enzymes. 2. In control anococcygeus muscles, DQ (10-100 microM) produced concentration-related inhibition (-log IC40 = 4.41) of relaxations to exogenous nitric oxide (NO; 15 microM). Nitrergic relaxations induced by field stimulation (10 Hz; 10 s train) were much less affected, 100 microM DQ reducing nitrergic relaxations by only 14 +/- 6%. 3. Following incubation with the Cu/Zn SOD inhibitor, diethyldithiocarbamate (DETCA; 3 mM; 45 min incubation; 10 min washout), the inhibitory effects of DQ on relaxations to NO were potentiated (-log IC40 = 5.22), and clear, concentration-related inhibitions of nitrergic relaxations were now observed (-log IC40 = 4.54). In both cases, these inhibitions were partially reversed by Cu/Zn SOD (250 u ml-1). In DETCA-treated tissues, DQ (100 microM) also reduced relaxations to sodium nitroprusside (1 microM) and S-nitroso-glutathione (30 microM), but potentiated those to 8-Br-cyclic GMP (100 microM). 4. Neither hydroquinone (HQ: 100 microM) nor 1,4-benzoquinone (BQ: 100 microM), both of which reduced responses to exogenous NO, inhibited relaxations induced by field stimulation in DETCA-treated tissues. Indeed, when added during DQ-induced inhibition of nitrergic relaxations, both HQ and BQ produced partial reversal of the block. 5. DQ had no effect on the detection of superoxide anions estimated via the xanthine:xanthine oxidase chemiluminescence assay, or of authentic NO as measured by a chemical microsensor. However, the detection of both superoxide anions and NO in these assays was inhibited by inclusion of either HQ or BQ. 6. The results support the proposal that nitrergic transmission in the peripheral nervous system is protected by Cu/Zn SOD activity in the region of the neuroeffector junction, and this may explain the lack of effect of superoxide anion generating drugs such as DQ. Such an explanation does not hold for either HQ or BQ, which appear to be acting directly as free radical scavengers in these experiments.
authors.affiliation: Biomedical Sciences Division, King's College London.
authors.forename: E
authors.initials: E
authors.lastname: Lilley
authors.forename: A
authors.initials: A
authors.lastname: Gibson
mesh_terms: D000818:Animals; D016227:Benzoquinones / Q000737:chemistry / Q000494:pharmacology*; D002614:Chelating Agents / Q000494:pharmacology; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000494:pharmacology; D004050:Ditiocarb / Q000494:pharmacology; D006873:Hydroquinones / Q000737:chemistry / Q000494:pharmacology; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D009126:Muscle Relaxation / Q000187:drug effects; D009130:Muscle, Smooth / Q000187:drug effects* / Q000502:physiology; D009569:Nitric Oxide / Q000037:antagonists & inhibitors* / Q000737:chemistry; D010084:Oxidation-Reduction; D013482:Superoxide Dismutase / Q000737:chemistry; D013481:Superoxides / Q000737:chemistry
chemical_list: D016227:Benzoquinones; D002614:Chelating Agents; D006873:Hydroquinones; D013481:Superoxides; C016276:8-bromocyclic GMP; D009569:Nitric Oxide; D004050:Ditiocarb; D013482:Superoxide Dismutase; D006152:Cyclic GMP; C034257:duroquinone; C031927:hydroquinone