Recent research has suggested that mild denervation of the neocortex of adult rats may facilitate neuronal growth in response to behavioral changes. Astrocytes react to denervation, produce growth-promoting factors and are a potential mediator of this denervation-facilitated growth. The present study assessed whether astrocytic reactions to denervation vary dependent upon post-injury behavioral experience. Denervation of the transcallosal afferents to the motor cortex was induced via partial transections of the corpus callosum. Transected- or sham-operated rats were then either forced to use the opposite forelimb (via limb-restricting vests) or permitted to use both forelimbs normally for 8 days. In the motor cortex, the surface density of glial fibrillary acidic protein (GFAP)-immunoreactive (IR) astrocytic processes and the density of basic fibroblast growth factor (FGF-2)-IR glial cells was significantly increased as a result of transections alone and as a result of forced forelimb-use alone in comparison to controls. The combination of transections and forced-use significantly enhanced GFAP-IR in comparison to all other groups, but did not further enhance FGF-2-IR. These findings are consistent with behavior and denervation having interactive influences on astrocytic reactivity in the motor cortex. These results also raise the possibility that astrocyte-mediated support of neural restructuring after brain injury might be enhanced with appropriate post-injury behavioral manipulations.