Neuropathy is a common complication of diabetes, reducing neuronal function, including impaired regeneration. Failure of CNS axons to regenerate after injury is at least partly due to intrinsic neuronal factors. Cyclic AMP (cAMP) activation promotes sensory axon regeneration through an Epac-mediated pathway. We therefore explored whether diabetic neurons are defective in cAMP-Epac signalling. We used cultured dissociated dorsal root ganglion (DRG) neurons from terminally-anaesthetised streptozotocin -induced diabetic and age-matched control rats to study their growth and responses to axon guidance cues. Growth of DRG neurons on a laminin substrate was significantly greater in controls compared with diabetic rats after 48 hours (35.9 ± 3.1% control neurons grew processes of at least 3 cell body diameters (3 cbd) vs. 27.4 ± 2.0% diabetic neurons, p<0.05; two-tailed, unpaired Student’s t-test was used in all statistical analyses). The cAMP agonist, Sp-cAMPS (20µM), enhanced neurite outgrowth of diabetic and control neurons at both 24 and 48 hours, but a selective Epac agonist, 8 pMeOPT-2′-O-Me-cAMP (2µM), significantly increased neurite outgrowth in control neurons (18.8 ± 1.6%) compared to diabetic neurons (12.6 ± 1.8%, p<0.05) after 24 hours. By 48 hours, the Epac agonist significantly improved the outgrowth of diabetic neurons (39.1 ± 1.5% Epac agonist vs. 27.4 ± 2.0% control, p<0.05), which was similar to the effect of the Epac agonist on age-matched control rat neurons at this time point (33.1 ± 3.2%). Activating cAMP or Epac also reversed the repulsive response of diabetic DRG neuronal growth cones to a gradient of the cAMP-dependent trophic factor, nerve growth factor (NGF). After 48 hours 13 out of 16 control growth cones turned towards the gradient, with a mean turning angle (MTA) of 17.6 ± 6.5°, while 9 out of 11 diabetic growth cones were repelled by a NGF gradient, showing a MTA of -22.5 ± 9.1°(p<0.001). Bath application of 20µM Sp-cAMPS, to elevate cAMP activity in diabetic neurons, converted diabetic growth cone repulsion to attraction at the 48 hour time point (MTA with Sp-cAMPS was 11.9 ± 4.9° vs. -22.5 ± 9.1° without Sp-cAMPS, p<0.001), as did bath application of the Epac agonist (MTA of 5.7 ± 5.9° with agonist vs -22.5 ± 9.1° without agonist, p<0.01). Blocking cAMP with the Rp-cAMPS isomer (20µM), which binds to Epac but does not activate it, abolished both the attraction of control growth cones in a NGF gradient and repulsion of diabetic neurons. An 18.9% decrease (p<0.05) in Epac expression was seen in diabetic neurons by Western blot and a 35% decrease (p<0.05) of Epac expression in growth cones by immunocytochemistry. Together, these results suggest that adult diabetic rat neurons exhibit reduced expression of Epac, resulting in a compromised ability to regenerate neurites and an inability to respond to cAMP-dependent chemoattractive guidance cues.