We assessed passive static stretching (PS) effects on the spatial distribution of muscle excitation (SDME) of the stretched (SL) and contralateral non-stretched limb (CL) during maximum voluntary isometric contractions (MVC). Before (PRE) and after 5-min PS, immediately (POST), at min 5 (POST5) and 10 (POST10), range of motion (ROM), maximal M-wave (Mmax) and MVC of both limbs were assessed in thirty men. During MVC, high-density surface electromyographic signals from the gastrocnemius medialis (GM) and lateralis (GL) were collected. The root mean square (RMS) and centroid coordinates were then obtained. During PS, discomfort perception (VAS), proximal and distal GM architecture were recorded. At POST, ROM increased and MVC decreased together with RMS in both limbs (P < 0.05). A cranio-caudal shift in SDME occurred in both muscles of SL and CL (P < 0.01) that persisted only in SL until POST5 in GM (P = 0.04), and POST10 in GL (P = 0.01). During PS, VAS was high (>8.0), and fascicle length and angle increased from rest (P < 0.01). No differences between GM portions were found in muscle architecture and in Mmax (P > 0.05). The results suggest involvement of central neural mechanisms in SDME shift. The prolonged effect in SL compared to CL indicates a possible additional contribution from mechanical mechanisms.

