SUMMARYMusculoskeletal models are often created by making detailed anatomical measurements of muscle properties. These measurements can then be used to determine the parameters of canonical models of muscle action. We describe here a complementary approach for developing and validating muscle models, using in situ measurements of muscle actions. We characterized the actions of two rat hindlimb muscles: the gracilis posticus (GRp) and the posterior head of biceps femoris (BFp; excluding the anterior head with vertebral origin). The GRp is a relatively simple muscle, with a circumscribed origin and insertion. The BFp is more complex, with an insertion distributed along the tibia. We measured the six-dimensional isometric forces and moments at the ankle evoked from stimulating each muscle at a range of limb configurations. The variation of forces and moments across the workspace provides a succinct characterization of muscle action. We then used this data to create a simple muscle model with a single point insertion and origin. The model parameters were optimized to best explain the observed force-moment data. This model explained the relatively simple muscle, GRp, very well (R 2 >0.85). Surprisingly, this simple model was also able to explain the action of the BFp, despite its greater complexity (R 2 >0.84). We then compared the actions observed here with those predicted using recently published anatomical measurements. Although the forces and moments predicted for the GRp were very similar to those observed here, the predictions for the BFp differed. These results show the potential utility of the approach described here for the development and refinement of musculoskeletal models based on in situ measurements of muscle actions.