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Even under isometric conditions, muscle contractions are associated with some degree of fiber shortening. The effects of muscle shortening on extracellular electromyographic potentials have not been characterized in detail. Moreover, the anatomical, biophysical, and detection factors influencing the muscle-shortening effects have been neither identified nor understood completely. Herein, we investigated the effects of muscle shortening on the amplitude and duration characteristics of single-fiber, motor unit, and compound muscle action potentials. We found that, at the single-fiber level, two main factors influenced the muscle-shortening effects: (1) the electrode position and distance relative to the myotendinous zone and (2) the electrode distance to the maxima of the dipole field arising from the stationary dipole created at the fiber-tendon junction. Besides, at the motor unit and muscle level, two additional factors were involved: (3) the overlapping between the propagating component of some fibers with the non-propagating component of other fibers and (4) the spatial spreading of the fiber-tendon junctions. The muscle-shortening effects depend critically on the electrode longitudinal distance to the myotendinous zone. When the electrode was placed far from the myotendinous zone, muscle shortening resulted in an enlargement and narrowing of the final (negative) phase of the potential, and this enlargement became less pronounced as the electrode approached the fiber endings. For electrode locations close to the myotendinous zone, muscle shortening caused a depression of both the main (positive) and final (negative) phases of the potential. Beyond the myotendinous zone, muscle shortening led to a decrease of the final (positive) phase. The present results provide reference information that will help to identify changes in MUPs and M waves due to muscle shortening, and thus to differentiate these changes from those caused by muscle fatigue. Graphical abstract

Effects of muscle shortening on single-fiber, motor unit, and compound muscle action potentials