Improved motor unit number estimate when motor unit alternation is addressed

Motor unit number estimation (MUNE) is important for determining motoneuron survival with age or in conditions such as amyotrophic lateral sclerosis or spinal cord injury. The original incremental method and approaches that were introduced to minimize alternation (e.g., multiple-point stimulation) are most commonly used, but one must accept the limitation that alternation of motor units may still inflate the estimate. Alternation occurs because axon thresholds are probabilistic and overlap for different axons; therefore, different combination of motor units may respond at a given stimulus intensity. Our aims were to quantify motor unit alternation systematically in the thenar muscles of 35 healthy adults by digital subtraction of EMG and force, and to compare MUNE with and without alternation. Alternation was prevalent, with one to nine occurrences in the first 7 to 11 steps in EMG in 34 of 35 muscles. It occurred in the first 3 steps in EMG in 49% of muscles. This alternation resulted in fewer units than steps in EMG (3 to 10 units at step 7 to 11). Accounting for alternation using digital subtraction reduced MUNE by up to 50%, day-to-day, and between-participant variability in MUNE. These results highlight the need to quantify alternation to improve the reliability and precision of motor unit number estimates, which will allow for detection of smaller changes in motoneuron survival with age, various health conditions, and/or due to an intervention. NEW & NOTEWORTHY Motor unit alternation was quantified systematically for the first time, addressing a major limitation of motor unit number estimates. Accounting for alternation decreased motor unit number estimates, and improved the reliability and precision of the motor unit number estimate, which will allow smaller, clinically relevant changes in motoneuron survival to be detected.