Opposing actions of neuronal nitric oxide synthase isoforms in formalin-induced pain in mice

The role of central and peripheral neuronal nitric oxide synthase (nNOS) splice variants in the development of inflammatory hyperalgesia was investigated using the formalin test. Supraspinal administration of the NOS inhibitor NOArg lowered both the first and second phase of the formalin response. An oligodeoxynucleotide targeting four nNOS isoforms given supraspinally also reduced the formalin response of both phases. Supraspinal antisense mapping suggested that this effect results from the nNOS-1 splice variant, implying that nNOS-1 is important in mediating formalin pain. At the spinal level, antisense mapping suggested a role of both the nNOS-1 and the nNOS-β variants in producing formalin pain. Conversely, an antisense selective against nNOS-2 had an opposing effect against the first phase, increasing its intensity. This result, which was similar to prior studies examining opioid actions, implies that endogenous nNOS-2 activity acted to minimize pain perception. Locally in the foot, arginine, the precursor for NO, increased the phase II response at low doses while higher doses reduced the response. This complex biphasic response suggested opposing NOS actions. Local antisense mapping again showed that nNOS-1 is involved in producing phase II of the formalin response while nNOS-2 had an opposite effect similar to that seen spinally. Finally, downregulation of nNOS-1 by antisense prevented tolerance to morphine in both the tail-flick and the formalin test. Together, these observations illustrate the complexity of nNOS in pain perception and the existence of opposing nNOS systems likely due to splice variants of nNOS.