1. Twenty-one days of bed rest alter motor unit properties and neuromuscular junction transmission in young adultsFabio Sarto, Miloš Kalc, Evgeniia Motanova, Martino V. Franchi, Daniel W Stashuk, Nina Murks, Giacomo Valli, Samuele Negro, Tomaž Prašnikar, Mladen Gasparini, Giovanni Martino, Giuseppe De Vito, Aleš Holobar, Boštjan Šimunič, Rado Pišot, Marco Vincenzo Narici, 2025, izvirni znanstveni članek Povzetek: Previous studies showed that properties of higher‐threshold motor units (MUs) and neuromuscular junction (NMJ) function are preserved during short‐term disuse. This study aimed to test how a longer disuse period affects MU properties, NMJ transmission, and NMJ morphology remodeling. Nine young healthy males (age: 18‐29 years) underwent 21 days of horizontal bed rest. Pre‐ (BR0) and post‐bed rest (BR21), quadriceps maximal voluntary contraction (MVC), and size were assessed. We combined intramuscular electromyography (iEMG) and high‐density surface electromyography (HDsEMG) recordings on the vastus lateralis to assess MU properties at 25% and 50% of MVC. Muscle biopsies and blood samples were also collected. Quadriceps MVC and size decreased at BR21. We found alterations in MU properties at both contraction intensities, including reduced discharge rate, MU potential area changes, and increased complexity. NMJ transmission was found to be reduced at BR21 at 25% MVC. This functional NMJ impairment was biochemically corroborated by an increase in serum C‐terminal agrin fragment concentration, a biomarker of NMJ instability. In addition, a direct assessment of NMJ morphology revealed the presence of some denervated NMJs exclusively at BR21. In conclusion, 21‐day bed rest altered MU properties across different contraction intensities and impaired NMJ transmission with initial signs of remodeling/denervation. Disuse duration appears to be a critical factor, as previous shorter studies failed to detect some of these changes. We believe these findings are clinically relevant for disuse after trauma, surgery, or illness, and may support the development of effective countermeasures.
Ključne besede: disuse, unloading, physical inactivity, electrophysiology, electromyography
Objavljeno v DiRROS: 13.05.2025; Ogledov: 590; Prenosov: 257
 Celotno besedilo (4,37 MB)
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2. Neuromuscular junction instability with inactivity : morphological and functional changes after 10 days of bed rest in older adultsStefano Schiaffino, Evgeniia Motanova, Fabio Sarto, Samuele Negro, Marco Pirazzini, Ornella Rossetto, Michela Rigoni, Daniel W Stashuk, Mladen Gasparini, Boštjan Šimunič, Rado Pišot, Marco Vincenzo Narici, 2025, izvirni znanstveni članek Povzetek: The neuromuscular junction (NMJ) plays a key role in modulating muscle contraction,but the impact of short-term disuse on NMJ structure and function, particularly in older humans,remains unclear. This study aimed to investigate NMJ alterations following 10 days of horizontalbed rest in 10 older males (68.5 ± 2.6 years). Before and after bed rest, vastus lateralis musclebiopsies were obtained to evaluate NMJ morphology, intramuscular EMG (iEMG) was recordedto assess NMJ function and blood samples were collected to determine circulating C-terminal agrinfragment (CAF) concentration, a biomarker of NMJ remodelling. In a sub-cohort of six participantswho had NMJs in both pre- and post-bed rest biopsies, we observed altered NMJ morphology,including reduced overlap between NMJ terminals, as well as increased endplate area and peri-meter. CAF concentration was elevated after bed rest, suggesting ongoing NMJ remodelling. iEMGanalysis showed increased motor unit potential complexity and reduced firing rate. In addition,we observed impaired NMJ transmission, inferred from increased near-fibre jiggle and segmentjitter. These findings suggest that older male individuals are susceptible to NMJ remodelling andimpaired transmission with short-term disuse, providing valuable insights into the morphologicaland functional consequences of inactivity in an ageing population. Our study highlights theimportance of developing interventions for mitigating the detrimental consequences of inactivityon neuromuscular health in older adults, which they frequently experience following injury, trauma,illness or surgery.
Ključne besede: ageing, C-terminal agrin, fragment disuse, electromyography, motor unit, NMJ unloading
Objavljeno v DiRROS: 19.03.2025; Ogledov: 697; Prenosov: 431
Celotno besedilo (2,13 MB)
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3. Initial glutathione depletion during short-term bed rest : pinpointing synthesis and degradation checkpoints in the γ-glutamyl cycleFilippo Giorgio Di Girolamo, Filippo Mearelli, Mariella Sturma, Nicola Fiotti, Kaja Teraž, Alja Ivetac, Alessio Nunnari, Pierandrea Vinci, Boštjan Šimunič, Rado Pišot, Gianni Biolo, 2024, izvirni znanstveni članek Povzetek: Hypokinesia triggers oxidative stress and accelerates the turnover of the glutathione system via the γ-glutamyl cycle. Our study aimed to identify the regulatory checkpoints controlling intracellular glutathione levels. We measured the intermediate substrates of the γ-glutamyl cycle in erythrocytes from 19 healthy young male volunteers before and during a 10-day experimental bed rest. Additionally, we tracked changes in glutathione levels and specific metabolite ratios up to 21 days of bed rest. Using gas chromatography-mass spectrometry and the internal standard technique, we observed a 9 ± 9% decrease in glutathione levels during the first 5 days of bed rest, followed by an 11 ± 9% increase from the 5th to the 10th day, nearly returning to baseline ambulatory levels. The cysteinyl-glycine-to-glutathione ratio, reflecting γ-glutamyl cyclotransferase activity (a key enzyme in glutathione breakdown), rose by 14 ± 22% in the first 5 days and then fell by 10 ± 14% over the subsequent 5 days, again approaching baseline levels. Additionally, the γ-glutamyl cysteine-to-cysteine ratio, indicative of γ-glutamyl cysteine synthetase activity (crucial for glutathione synthesis), increased by 12 ± 30% on day 5 and by 29 ± 41% on day 10 of bed rest. The results observed on day 21 of bed rest confirm those seen on day 10. By calculating the ratio of product concentration to precursor concentration, we assessed the efficiency of these key enzymes in glutathione turnover. These results were corroborated by directly measuring glutathione synthesis and degradation rates in vivo using stable isotope techniques. Our findings reveal significant changes in glutathione kinetics during the initial days of bed rest and identify potential therapeutic targets for maintaining glutathione levels.
Ključne besede: antioxidant, muscle unloading, glutathion turnover, γ-glutamyl cycle, gamma-glutamyl cycle
Objavljeno v DiRROS: 21.11.2024; Ogledov: 760; Prenosov: 1549
Celotno besedilo (1,84 MB)
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4. Neuromuscular junction instability and altered intracellular calcium handling as early determinants of force loss during unloading in humansElena Monti, Carlo Reggiani, Martino V. Franchi, Luana Toniolo, Marco Sandri, Andrea Armani, Sandra Zampieri, Boštjan Šimunič, Rado Pišot, Marco Vincenzo Narici, 2021, izvirni znanstveni članek Povzetek: Unloading induces rapid skeletal muscle atrophy and functional decline. Importantly, force is lost at a much higher rate than muscle mass. We aimed to investigate the early determinants of the disproportionate loss of force compared to that of muscle mass in response to unloading. Ten young participants underwent 10 days of bed rest (BR). At baseline (BR0) and at 10 days (BR10), quadriceps femoris (QF) volume (VOL) and isometric maximum voluntary contraction (MVC) were assessed. At BR0 and BR10 blood samples and biopsies of vastus lateralis (VL) muscle were collected. Neuromuscular junction (NMJ) stability and myofibre innervation status were assessed, together with single fibre mechanical properties and sarcoplasmic reticulum (SR) calcium handling. From BR0 to BR10, QFVOL and MVC decreased by 5.2% (P = 0.003) and 14.3% (P < 0.001), respectively. Initial and partial denervation was detected from increased neural cell adhesion molecule (NCAM)-positive myofibres at BR10 compared with BR0 (+3.4%, P = 0.016). NMJ instability was further inferred from increased C-terminal agrin fragment concentration in serum (+19.2% at BR10, P = 0.031). Fast fibre cross-sectional area (CSA) showed a trend to decrease by 15% (P = 0.055) at BR10, while single fibre maximal tension (force/CSA) was unchanged. However, at BR10 SR Ca2+ release in response to caffeine decreased by 35.1% (P < 0.002) and 30.2% (P < 0.001) in fast and slow fibres, respectively, pointing to an impaired excitation%contraction coupling. These findings support the view that the early onset of NMJ instability and impairment in SR function are eligible mechanisms contributing to the greater decline in muscle force than in muscle size during unloading.
Ključne besede: Ca2+ dynamics, muscle atrophy, neuromuscular junction instability, sarcoplasmic reticulum, single fibre atrophy, single fibre contractile impairment, unloading
Objavljeno v DiRROS: 16.06.2021; Ogledov: 2285; Prenosov: 2032
Celotno besedilo (3,39 MB)
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