Effects of short-term unloading and active recovery on human motor unit properties, neuromuscular junction transmission and transcriptomic profile

Sarto, F ORCID: https://orcid.org/0000-0001-8572-5147, Stashuk, DW ORCID: https://orcid.org/0000-0002-8072-8135, Franchi, MV ORCID: https://orcid.org/0000-0003-3165-4536, Monti, E ORCID: https://orcid.org/0000-0002-3767-0855, Zampieri, S ORCID: https://orcid.org/0000-0001-6970-0011, Valli, G, Sirago, G ORCID: https://orcid.org/0000-0001-7005-8708, Candia, J ORCID: https://orcid.org/0000-0001-5793-8989, Hartnell, LM, Paganini, M, McPhee, JS ORCID: https://orcid.org/0000-0002-3659-0773, De Vito, G, Ferrucci, L, Reggiani, C ORCID: https://orcid.org/0000-0001-8080-361X and Narici, MV ORCID: https://orcid.org/0000-0003-0167-1845 (2022) Effects of short-term unloading and active recovery on human motor unit properties, neuromuscular junction transmission and transcriptomic profile. Journal of Physiology, 600 (21). pp. 4731-4751. ISSN 0022-3751

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Abstract

Electrophysiological alterations of the neuromuscular junction (NMJ) and motor unit potential (MUP) with unloading are poorly studied. We aimed to investigate these aspects and the underlying molecular mechanisms with short-term unloading and active recovery (AR). Eleven healthy males underwent a 10-day unilateral lower limb suspension (ULLS) period, followed by 21-day AR based on resistance exercise. Quadriceps femoris (QF) cross-sectional area (CSA) and isometric maximum voluntary contraction (MVC) were evaluated. Intramuscular electromyographic recordings were obtained during 10% and 25% MVC isometric contractions from the vastus lateralis (VL). Biomarkers of NMJ molecular instability (serum c-terminal agrin fragment, CAF), axonal damage (neurofilament light chain) and denervation status were assessed from blood samples and VL biopsies. NMJ and ion channel transcriptomic profiles were investigated by RNA-sequencing. QF CSA and MVC decreased with ULLS. Increased CAF and altered NMJ transcriptome with unloading suggested the emergence of NMJ molecular instability, which was not associated with impaired NMJ transmission stability. Instead, increased MUP complexity and decreased motor unit firing rates were found after ULLS. Downregulation of ion channel gene expression was found together with increased neurofilament light chain concentration and partial denervation. The AR period restored most of these neuromuscular alterations. In conclusion, the human NMJ is destabilized at the molecular level but shows functional resilience to a 10-day unloading period at least at relatively low contraction intensities. However, MUP properties are altered by ULLS, possibly due to alterations in ion channel dynamics and initial axonal damage and denervation. These changes are fully reversed by 21 days of AR. (Figure presented.). Key points: We used integrative electrophysiological and molecular approaches to comprehensively investigate changes in neuromuscular integrity and function after a 10-day unilateral lower limb suspension (ULLS), followed by 21 days of active recovery in young healthy men, with a particular focus on neuromuscular junction (NMJ) and motor unit potential (MUP) properties alterations. After 10-day ULLS, we found significant NMJ molecular alterations in the absence of NMJ transmission stability impairment. These findings suggest that the human NMJ is functionally resilient against insults and stresses induced by short-term disuse at least at relatively low contraction intensities, at which low-threshold, slow-type motor units are recruited. Intramuscular electromyography analysis revealed that unloading caused increased MUP complexity and decreased motor unit firing rates, and these alterations could be related to the observed changes in skeletal muscle ion channel pool and initial and partial signs of fibre denervation and axonal damage. The active recovery period restored these neuromuscular changes.