Superoxide anions, free-radical scavengers, and nitrergic neurotransmission

1. There is now strong evidence that the L-arginine/nitric oxide (NO) pathway generates the transmitter released from certain nonadrenergic, noncholinergic nerves that mediate smooth-muscle relaxation in the respiratory, gastrointestinal, and urogenital tracts. In particular, nitric oxide synthase (NOS) has been detected in these nitrergic nerves, and nerve-induced relaxation can be prevented by NOS inhibitors. Thus, free-radical NO has been considered the putative transmitter candidate. 2. Despite such evidence, a number of superoxide anion-generating compounds and direct NO scavengers have been found to abolish relaxations to exogenous NO, but to have very little effect on relaxations in response to nitrergic field stimulation. A number of hypotheses have been put forward to explain this paradox: first, that the NO generated within the nerve is attached to a carrier molecule (such as a thiol) to form an adduct, that is released into the junctional gap and that is resistant to superoxide anions and other scavengers; second, that over short distances (up to 200 microns) the rapid diffusion characteristics of NO render it resistant to inhibition by scavengers; third, that NO is indeed released as a free radical, but that it is protected from radical scavengers by other substances present in the junctional region. 3. Recent experimental evidence supports the third explanation, because nitrergic relaxations, normally resistant to inhibition by superoxide anions, become sensitive following inactivation of copper/ zinc superoxide dismutase (Cu/Zn SOD); the inhibition can be reversed by adding exogenous Cu/Zn SOD (or ascorbate). In addition, the ability of two NO-scavenger compounds, hydroquinone and carboxy-PTIO, to inhibit relaxations to exogenous NO is prevented by certain physiological antioxidants (ascorbate and reduced glutathione in the case of hydroquinone, and ascorbate and alpha-tocopherol in the case of carboxy-PTIO). 4. Thus, it is possible that the presence of integrated antioxidant mechanisms within the tissue protects neuronally- released NO from attack by scavenging molecules; exogenous NO would be vulnerable before reaching the protection of the tissue, thus explaining the paradoxical effects mentioned. Organ antioxidant status may therefore be very important in preserving the potency of nitrergic transmission and in preventing NO from reacting with other compounds to produce cytotoxic metabolites (eg., with superoxide anions to form peroxynitrite).