The purpose of the present study is to investigate the origin of mechanical interactions between the rat extensor digitorum longus (EDL) muscle and the grouped tibialis anterior and extensor hallucis longus muscles (TA+EHL). The proximal and distal tendons of EDL as well as the tied distal tendons of TA+EHL were transected and connected to force transducers. Connective tissues at the muscle bellies of the anterior crural compartment were left intact. Supramaximal stimulation of the common peroneal nerve activated all muscles maximally and simultaneously. Length-isometric force characteristics of distal TA+EHL were assessed. Simultaneously, forces exerted at the proximal and distal tendons of EDL, kept at constant muscle-tendon complex length and position, were measured. Intermuscular interaction was tested in two conditions: (a) after full longitudinal compartmental fasciotomy, and (b) after blunt dissection of the intermuscular connective tissue linkages between EDL and TA+EHL. Note that in the latter condition, intermuscular myofascial pathways were eliminated. In the initial condition, lengthening TA+EHL by 12 mm increased proximal (by 0.14 N, i.e. 9.5%) and decreased distal EDL force (by 0.21 N, i.e. 11.8%), despite the fact that EDL muscle-tendon complex length was kept constant. Blunt dissection decreased TA+EHL and distal EDL forces at low TA+EHL lengths only, while proximal EDL force decreased for all TA+EHL lengths tested. The dissection caused no changes in the TA+EHL length effects on proximal EDL force. In contrast, the amplitude of change in the distal EDL force curve decreased significantly (by 39%) subsequent to blunt dissection. It is concluded that mechanical interaction between synergists originates from both intermuscular as well as extramuscular connective tissues. The highest contribution, however, should be ascribed to the extramuscular pathway. Copyright © 2005 S. Karger AG.