This study was set to investigate whether motor effects of nociceptin/orphanin FQ (N/OFQ) can be related to changes in primary motor cortex output. N/OFQ injected i.c.v. biphasically modulated motor performance, low doses being facilitating and higher ones inhibitory. These effects were counteracted by the N/OFQ receptor antagonist [Nphe(1) Arg(14),Lys(15)]N/OFQ-NH(2) (UFP-101) confirming the specificity of N/OFQ action. However, UFP-101 alone facilitated motor performance, suggesting that endogenous N/OFQ inhibits motor function. N/OFQ and UFP-101 injected into the substantia nigra reticulata but not motor cortex replicated these effects, suggesting motor responses were mediated by subcortical circuits involving the basal ganglia. Intracortical microstimulation technique showed that i.c.v. N/OFQ also biphasically modulated motor cortex excitability and movement representation. Low N/OFQ doses caused a leftward shift of threshold distribution curve in the forelimb area without affecting the number of effective sites. Conversely, high N/OFQ doses increased unresponsive and reduced excitable (movement) sites in vibrissa but not forelimb area. However, increased threshold currents and rightward shift of threshold distribution curve were observed in both areas, suggesting an overall inhibitory effect on cortical motor output. UFP-101 alone evoked effects similar to low N/OFQ doses, suggesting tonic inhibitory control over forelimb movement by endogenous N/OFQ. As shown in behavioral experiments, these effects were replicated by intranigral, but not intracortical, N/OFQ or UFP-101 injections. We conclude that N/OFQ receptors located in the substantia nigra reticulata mediate N/OFQ biphasic control over motor behavior, possibly through changes of primary motor cortex output.