Tibial nerves from control, untreated alloxan diabetic, and 4-week insulin treated alloxan diabetic rats were examined with light microscopy and computerized morphometric analysis of axons. Teased fiber preparations and electron microscopy were utilized to evaluate nerve degeneration. The insulin treatment regimens included daily injections of protamine zinc insulin (PZI), daily injections of ultralente insulin, and continuously delivered insulin through osmotic minipumps. Evaluation of axon:myelin ratios, teased fiber profiles, and electron microscopic cross sections of nerves demonstrated different degrees of neuropathic changes within the treated groups. The control group and untreated diabetic group showed little or no degeneration, while all insulin-treated groups showed evidence of Wallerian degeneration. Among these insulin treated groups, the PZI-treated group showed the greatest number of degenerating profiles while the minipump group showed the least. These data suggest that insulin treatment of alloxan diabetes results in axonal degeneration which closely resembles findings in human diabetic neuropathies. The substantially diminished number of degenerating axons seen in the osmotic minipump insulin-treated rats suggests that continuous delivery of insulin may decrease the neuropathic changes seen with single injection insulin therapy. Since virtually all insulin-dependent diabetic patients receive daily administration of insulin, the possibility that peripheral neuropathies may in part result from the insulin treatment requires more extensive investigation in a variety of animal models to separate the neuropathic effects of diabetes from the neuropathic effects of insulin therapy.