Hindlimb suspension results in rapid losses of muscle mass, which may in part, be explained by attenuated rates of muscle protein synthesis. Recently, reduced activity of mammalian target of rapamycin (mTOR) signaling has been implicated as a potential mediator of the decrement in muscle mass with hindlimb suspension. The purpose of this study was to examine the effect of acute resistance exercise, a known stimulant of muscle hypertrophy, on rates of protein synthesis with and without rapamycin, an inhibitor of mTOR, after only 4 days of hindlimb suspension in mature, male, Sprague-Dawley rats. Resistance exercise consisted of 2 sets of 25 repetitions using flywheel technology on the second and fourth day of hindlimb suspension. Muscles were obtained from rats 16 h after the last exercise bout and assessed for rates of muscle protein synthesis using the incorporation of radioactive phenylalanine. Isolated soleus muscles were incubated, in vitro, with and without insulin (a signaling agonist) and/or rapamycin (an mTOR inhibitor). Results demonstrate that soleus muscle mass was significantly reduced (p<0.05) with 4 days of hindlimb suspension (HS) but this loss of mass was not observed (p>0.05) with the addition of resistance exercise (HSRE). Rates of soleus muscle protein synthesis were diminished (p<0.05) after 4 days of HS, with or without insulin. HSRE also had reduced rates of muscle protein synthesis without insulin; however, acute insulin administration yielded higher (p<0.05) rates of synthesis in HSRE compared to HS or control, with or without insulin. Rapamycin diminished rates of synthesis in all groups (p<0.05), but insulin rescued rates of synthesis in both HS and HSRE to levels similar to insulin alone for each group, respectively, suggesting that alternate signaling pathways develop to increase protein synthesis in response to hindlimb suspension. These results demonstrate that the capacity for an augmented anabolic response to resistance exercise is maintained even after 4 days of hindlimb suspension, and that this response is independent of a rapamycin-sensitive pathway.