Measurement of forces of mono- and bi-articular muscles of an entire intact muscle compartment can allow for a comprehensive assessment of the effects of Botulinum toxin type A (BTX-A) both at and beyond the injection site, and in conditions close to those in vivo. The goal was to test the hypotheses that BTX-A affects (1) the forces of not only the injected but also the noninjected muscles of the compartment, and (2) epimuscular myofascial force transmission (EMFT). Two groups of Wistar rats were tested: Control (no BTX-A injected) and BTX (0.1 units of BTX-A were injected exclusively to the mid-belly of TA). Isometric forces were measured simultaneously at the distal tendons of the tibialis anterior (TA) at different lengths, the restrained extensor digitorum longus (EDL) and the extensor hallucis longus (EHL) muscles and at the proximal tendon of EDL. Five days post-injection, BTX-A did affect the total forces of all muscles significantly: (1) The TA force decreased differentially (by 46.6%-55.9%) for most lengths such that a significant negative correlation was found between force reductions and increased muscle length. The maximum TA force decreased by 47.3%. However, the muscle's length range of force production did not change significantly. (2) Distal and proximal EDL forces decreased (on average by 67.8% and 62.9%, respectively). (3) The EHL force also decreased (on average by 9.2%). The passive forces of only the TA showed a significant increase at higher lengths. EMFT effects were shown for the control group: (1) at the shortest TA lengths, the EDL proximo-distal force differences were in favor of the distal force, which was reversed at higher lengths. (2) the EHL force measured at the shortest TA length decreased (by 34%) as a function of TA lengthening. After BTX-A exposure, such EMFT effects disappeared for the EDL, whereas they remained as profound for the EHL. Exposure to BTX-A does affect forces of all muscles operating in an intact compartment. For the BTX-A injected muscle, the reduction in muscle force becomes less pronounced at higher muscle lengths. BTX-A also has effects on EMFT, however, these effects are not uniform within the anterior crural compartment. Decreased forces of the noninjected synergistic muscles suggest the presence of unintended additional effects of BTX-A both for the targeted distal joint and for the nontargeted proximal joint.

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