Chronic central nervous system demyelinating diseases result in long-term disability because of limited remyelination capacity and cumulative damage to axons. Corpus callosum demyelination in mice fed cuprizone provides a reproducible model of chronic demyelination in which the demyelinating agent can be removed to test modifications that promote recovery and to develop noninvasive neuroimaging techniques for monitoring changes in myelin and axons. We used the cuprizone model in mice with genetic deletion of fibroblast growth factor 2 (Fgf2) to determine the impact of FGF2 on axon pathology and remyelination after chronic demyelination. We also evaluated the ability of quantitative magnetic resonance diffusion tensor imaging (DTI) to distinguish the corresponding pathological changes in axons and myelin during the progression of demyelination and remyelination. During the recovery period after chronic demyelination, Fgf2-null mice exhibited enhanced remyelination that was detected using DTI measures of radial diffusivity and confirmed by electron microscopic analysis of the proportion of remyelinated axons. Ultrastructural analysis also demonstrated reduced axonal atrophy in chronically demyelinated Fgf2-null versus wild-type mice. This difference in axon atrophy was further demonstrated as reduced immunohistochemical detection of neurofilament dephosphorylation in Fgf2-null mice. Diffusion tensor imaging axial and radial diffusivity measures did not differentiate Fgf2-null mice from wild-type mice to correlate with changes in axonal atrophy during chronic demyelination. Overall, these findings demonstrate that attenuation of FGF2 signaling promotes neuroprotection of axons and remyelination, suggesting that FGF2 is an important negative regulator of recovery after chronic demyelination.