Although the Achilles tendon (AT) has been studied for more than a century, a complete understanding of the mechanical and functional consequences of AT structural organization is currently lacking. The aim of this study was to assess how joint angle configuration affects subtendon displacement and strain of soleus (SOL) and lateral gastrocnemius (LG) muscles. Knots sutured onto SOL and LG subtendons of 12 Wistar rats, were videotaped to quantify displacements and the ankle torque was assessed for different isometric activation conditions (i.e., individual and simultaneous) of the triceps surae muscles. Changing ankle and knee joint angle affected the magnitude of displacement, relative displacement and strain of both SOL and LG subtendons. SOL subtendon behavior was not only affected by changes in ankle angle, but also by changes in knee angle. Displacement of SOL subtendon decreased (28-49%), but strain increased in response to knee extension. Independent of joint angle configuration, stimulation of any combination of the muscles typically resulted in displacements and strains of LG and SOL subtendons. Typically SOL displaced more but LG displaced more when stimulated at longer muscle lengths. Our results demonstrate that the distinct subtendons of the Achilles tendon can move and deform differently, but are not fully independent. Within the AT, there appears to be a precarious balance between sliding allowance and mechanical connectivity between subtendons.