The present study was designed to determine the contribution of weight bearing to the adaptations of the plantaris (PL) to synergist removal. PL from female rats were exposed to 28 days of a simultaneous condition of synergist ablation and hindlimb suspension. At 28 days, contractile responses and morphological measures were obtained and compared with muscles that either had synergists intact or were weight bearing or a combination of both. Synergist ablation prolonged PL maximum isometric twitch tension (Pt), time to peak tension (12%), and one-half relaxation time (12%); increased Pt (26%), maximum isometric tetanic tension (Po, 44%), fatigue resistance (FI, 42%), and fast fiber cross-sectional area (FT CSA, 20%); and decreased Pt/Po (13%) over nonablation counterparts. Suspension decreased PL Pt (26%), Po (26%), rest length (16%), FT CSA (31%), and slow-twitch fiber (ST) number (24%) but increased FI (75%) over weight-bearing counterparts. PL from weight-bearing animals were heavier than from suspended animals, and the extent of this response was greatest after synergist removal. Whole muscle and ST CSA and ST area contribution were greater only in weight-bearing synergist ablation muscles. Daily weight bearing (4 h) in synergist ablation hindlimb suspension groups caused PL weights and ST expressions to be halfway between 24-h suspension and 24-h weight-bearing groups. Our results suggest that weight bearing is not essential to the induction of several adaptations associated with synergist ablation but is required to cause the large muscle mass and ST expression characteristic of this model.