Nitric oxide synthase-1 (NOS1) is involved in several forms of plasticity including hippocampal-dependent learning and memory, experience-dependent plasticity in the barrel cortex, and long-term potentiation (LTP) in the hippocampus and neocortex. NOS1 also contributes to ischemic damage during stroke and has a stronger deleterious effect in males than females. We therefore investigated whether the role of NOS1 in plasticity might also be sex specific. We tested LTP in the layer IV-II/III pathway between barrel columns and experience-dependent plasticity in the barrel cortex of αNOS1 knock-out mice and their wild-type littermates. We found that LTP was absent in male αNOS1 knock-out mice but not in females and that the residual LTP in females was not NO dependent. We also found that experience-dependent potentiation due to single whisker experience was significantly reduced in male αNOS1 knockouts but was unaffected in females. The αNOS1 knockout had a small effect on the development of the barrels, which were reduced in size by 20% compared with wild types, but this effect was not sex specific. We therefore conclude that neocortical plasticity mechanisms differ between males and females at the synaptic level, either in their basic plasticity induction pathways or in their ability to compensate for loss of αNOS1.