Differential nitric oxide synthase activity, cofactor availability and cGMP accumulation in the central nervous system during anaesthesia

We investigated the effects of anaesthesia on dynamic nitric oxide production, concentrations of tetrahydrobiopterin and the accumulation of cyclic GMP (cGMP) in the rat central nervous system (CNS). Rats were assigned to anaesthesia with halothane, isoflurane, pentobarbital, diazepam, ketamine or xenon (n=6 per group). After 30 min, [14C]l-arginine (i.v.) was given and, after a further 60 min of anaesthesia, rats were killed and exposed immediately to focused microwave radiation. After removal of the brain and spinal cord, nitric oxide production from radiolabelled arginine (and hence nitric oxide synthase activity during anaesthesia) was measured as [14C]l-citrulline by scintillation counting. cGMP was determined by enzyme immunoassay and tetrahydrobiopterin by fluorescence HPLC, in brain regions and the spinal cord. Nitric oxide synthase activity was similar in all brain regions but was lower in the spinal cord, and was unaffected by anaesthesia. cGMP was similar in all areas of the CNS and was significantly decreased in rats anaesthetized with halothane. Isoflurane produced similar effects. In contrast, ketamine and xenon anaesthesia increased cGMP in the spinal cord, brainstem and hippocampus. Diazepam and pentobarbital had no effect. Tetrahydrobiopterin concentrations were similar in all areas of the CNS and were increased in the cortex and hippocampus after anaesthesia. We have shown profound differential effects of anaesthesia on the nitric oxide pathway in the rat CNS.