Cyclic adenosine monophosphate (cAMP) has been intensively studied in recent years in order to elucidate its contribution in intracellular signalling mechanisms that regulate axon growth and guidance, and also to test if its activation can promote axon regeneration after injury. Cyclic guanosine monophosphate (cGMP), however, has been given considerably less attention even though it too mediates intracellular signalling cascades activated by extracellular guidance cues. cGMP can promote neurite outgrowth in neuronal cell lines but its role in promoting growth and regeneration of primary neurons is not well established. Here, we have examined the effects of elevating cGMP activity on axon growth, guidance and regeneration in vitro. We have found that, like cAMP elevation, activation of cGMP increases rat dorsal root ganglion (DRG) neurite outgrowth on a polylysine substrate and that asymmetric cGMP elevation promotes attractive growth cone turning. When grown in an in vitro model of axon regeneration activation of cGMP alone was not sufficient to promote adult neurite outgrowth. However, when combined with cAMP elevation substantial regeneration of adult neurites is achieved, superior to that achieved with either cAMP or cGMP alone. Regeneration is enhanced still further with simultaneous application of a Nogo receptor blocking peptide, suggesting this combinatorial strategy could achieve far greater axon regeneration in vivo than targeting individual cell signalling mechanisms. (C) 2009 Elsevier B.V. All rights reserved.