Biology subjects

Atherton, P. J.

Publications and source records attributed to Atherton, P. J..

11 recordsLinked to original sources

bioRxiv2025

Proteostasis Modelling using Deuterated Water Metabolic Labeling and Data-Independent Acquisition Mass Spectrometry

We present the first application of a deuterated water metabolic labeling workflow coupled with data-independent acquisition (DIA) tandem mass spectrometry (MS/MS) for quantifying label enrichment in MS/MS to study protein turnover. The approach automates the turnover rate determination from combined precursor and fragment ions. The truncation of the observed isotope distributions of fragments is overcome by implementing an approach to determining the label enrichment from two mass isotopomers. The high redundancy of fragment ions provides a confident assessment for deuterium enrichment. The DIA approach provides increased proteome coverage and depth for quantifying protein turnover rates compared to the traditional data-dependent (DDA) approach. This novel approach is validated in a murine myotube model of muscle hypertrophy and atrophy through the treatment with insulin-like growth factor 1 (IGF-1) and dexamethasone (Dex), respectively.

bioinformatics↗

bioRxiv2025

Task Complexity and Limb Dominance Modulate the Effects of Ageing on Neuromuscular Function

BackgroundNeuromuscular function is critical for independence in ageing, yet asymmetries between dominant and non-dominant limbs, arising from central or peripheral mechanisms, are not well understood. This study examined age- and limb-related differences and motor unit (MU) firing behaviour of the vastus lateralis under tasks of varying difficulty. MethodsTwenty-one young (22 {+/-} 4 years; 15M, 6F) and seventeen older adults (74 {+/-} 5 years; 12M, 5F) performed constant and variable force unilateral isometric knee extensions. In both limbs, high-density surface electromyography signals were decomposed into MU spike trains. Force control and MU firing properties were analysed using multilevel mixed-effects regression models. ResultsOlder adults showed reduced maximal muscle strength (p<0.001) and increased force tracking error (p=0.008), MU firing rate (MUFR) was significantly lower in older adults during constant contractions (p = 0.001) and trended toward lower during variable contractions (p=0.061). MUFR variability showed a significant Leg x AgeGroup interaction (p<0.001); older adults had greater variability in non-dominant legs, while younger adults showed the opposite. With variable force contractions in both age groups, MUFR was higher during ascending segments with greater variability during descending segments. ConclusionNeuromuscular ageing involves asymmetric adaptations rather than a uniform decline, with leg dominance effects become more pronounced under variable force modulation. Task difficulty amplifies these asymmetries, underscoring the need to consider limb-specific neural control in age-related motor assessments.

physiology↗

bioRxiv2025

Rapamycin exerts its geroprotective effects in the ageing human immune system by enhancing resilience against DNA damage

mTOR inhibitors such as rapamycin are among the most robust life-extending interventions known, yet the mechanisms underlying their geroprotective effects in humans remain incompletely understood. At non-immunosuppressive doses, these drugs are senomorphic, i.e. they mitigate cellular senescence, but whether they protect genome stability itself has been unclear. Given that DNA damage is a major driver of immune ageing, and immune decline accelerates whole-organism ageing, we tested whether mTOR inhibition enhances genome stability. In human T cells exposed to acute genotoxic stress, we found that rapamycin and other mTOR inhibitors suppressed senescence not by slowing protein synthesis, halting cell division, or stimulating autophagy, but by directly reducing DNA lesional burden and improving cell survival. Ex-vivo analysis of aged immune cells from healthy donors revealed a stark enrichment of markers for DNA damage, senescence, and mTORC hyperactivation, suggesting that human immune ageing may be amenable to intervention by low-dose mTOR inhibition. To test this in vivo, we conducted a placebo-controlled experimental medicine trial in older adults administered with low-dose rapamycin. p21, a marker of DNA damage-induced senescence, was significantly reduced in immune cells from the rapamycin compared to placebo group. These findings reveal a previously unrecognised role for mTOR inhibition: direct genoprotection. This mechanism may help explain rapamycins exceptional geroprotective profile and opens new avenues for its use in contexts where genome instability drives pathology, ranging from healthy ageing, clinical radiation exposure, and even the hazards of cosmic radiation in space travel.

immunology↗

bioRxiv2024

Theta-gamma transcranial alternating current stimulation enhances motor skill acquisition in healthy young and older adults

Theta-gamma transcranial alternating current stimulation (TG tACS) over primary motor cortex (M1) can improve motor skill acquisition in young adults, but the effect on older adults is unknown. This study investigated the effects of TG tACS on motor skill acquisition and M1 excitability in 18 young and 18 older adults. High-definition TG tACS (6 Hz theta, 75 Hz gamma) or sham tACS was applied over right M1 for 20 minutes during a ballistic left-thumb abduction motor training task performed in two experimental sessions. Motor skill acquisition was quantified as changes in movement acceleration during and up to 60 minutes after training. Transcranial magnetic stimulation (TMS) was used to assess changes in M1 excitability with motor-evoked potentials (MEP) and short-interval intracortical inhibition (SICI) before and after training. We found that TG tACS increased motor skill acquisition compared with sham tACS in young and older adults (P < 0.001), with greater effects for young adults (P = 0.01). The improved motor performance with TG tACS lasted at least 60 minutes after training in both age groups. Motor training was accompanied by greater MEP amplitudes with TG tACS compared to sham tACS in young and older adults (P < 0.001), but SICI did not vary between tACS sessions (P = 0.40). These findings indicate that TG tACS over M1 improves motor skill acquisition and alters training-induced changes in M1 excitability in healthy young and older adults. TG tACS may therefore be beneficial to alleviate motor deficits in the ageing population. Key Points SummaryO_LITheta-gamma transcranial alternating current stimulation (TG tACS) can improve motor function in healthy young adults, but the effect on older adults is unknown. C_LIO_LIWe found that TG tACS improved motor skill acquisition with long-lasting effects in healthy young and older adults, but effects were stronger in young adults. C_LIO_LITranscranial magnetic stimulation showed that TG tACS altered the training-induced changes in motor cortex excitability, but there was no effect of TG tACS on intracortical inhibition in young or older adults. C_LIO_LIOur data suggest that TG tACS represents a promising approach to improve motor skill acquisition throughout the lifespan, and may be beneficial in older patient populations that experience motor or cognitive deficits. C_LI

neuroscience↗

bioRxiv2023

Menstrual cycle associated alteration of vastus lateralis motor unit function

Estrogen and progesterone are the primary female sex hormones and have net excitatory and inhibitory effects, respectively, on neuronal function. Fluctuating concentrations across the menstrual cycle has led to several lines of research in relation to neuromuscular function, yet evidence from animal and cell culture models have yet to be demonstrated in human motor units (MU) coupled with quantification of circulating hormones. Intramuscular electromyography (iEMG) was applied to record MU potentials (MUP) and corresponding MUP trains (MUPT) from the vastus lateralis of eumenorrheic females during the early follicular, ovulation and mid luteal phases of the menstrual cycle, alongside assessments of neuromuscular performance. Multi-level regression models were applied to explore effects of time and of contraction level. Statistical significance was accepted as p<0.05. Knee extensor maximum voluntary contraction (MVC), jump power, force steadiness, and balance did not differ across the menstrual phases (all p>0.4). Firing rate of low threshold MU (10% MVC) was reduced during phases of high progesterone ({beta}=-0.82Hz, p<0.001), with no difference in MUPs analysed from 25% MVC contractions. MUPs were more complex during ovulation and mid luteal phase (p<0.03), with no change in neuromuscular junction transmission instability (p>0.3). Assessments of neuromuscular performance did not differ across the menstrual cycle. The suppression of low threshold MU firing rate during periods of increased progesterone may suggest a potential inhibitory effect and an alteration of recruitment strategy, however this had no discernible effect on performance. These findings highlight contraction level dependent modulation of VL MU function over the eumenorrheic cycle.

physiology↗

bioRxiv2023

Reduced motoneuron excitability and common synaptic inputs of the aged human vastus lateralis

Although muscle atrophy may partially account for age-related strength decline, it is further influenced by alterations of neural input to muscle. Persistent inward currents and the level of common synaptic inputs to motoneurons influence neuromuscular function. However, these have not yet been described in aged human quadriceps. High density surface electromyography (HDsEMG) signals were collected from the vastus lateralis of 15 young (mean{+/-}SD, 23 {+/-} 5 y) and 15 older (67 {+/-} 9 y) men during submaximal sustained and 20-s ramped contractions. HDsEMG signals were decomposed to identify individual motor unit discharges, from which delta F and intramuscular coherence were estimated. Older participants produced significantly lower knee extensor torque (p<0.001) and poorer force tracking ability (p<0.001) than young. Older participants also had lower delta F (p=0.001) and coherence estimates in the alpha frequency band (p<0.001) during ramp contractions when compared to young. Persistent inward currents and common synaptic inputs are lower in the vastus lateralis of older males when compared to young. These data highlight altered neural input to the clinically and functionally important quadriceps, further underpinning age-related loss of function which may occur independently of the loss of muscle mass. Key PointsO_LIThe age-related loss of muscle mass is exceeded by the loss of function, which is influenced by structural and functional alterations of the nervous system. C_LIO_LIMotoneuronal persistent inward currents and common synaptic inputs play an important role in the activation of motor units and subsequent force generation and control ability. C_LIO_LIHere we show reduced estimates of persistent inward currents and lower common synaptic inputs to older vastus lateralis, potentially contributing to observed lower strength and poorer force tracking. C_LIO_LIThese findings highlight decrements of the aged human motor system, accompanied by muscle atrophy in functionally relevant muscle groups, which should be considered in the application of interventions targeting aged human muscle. C_LI

physiology↗

bioRxiv2022

Motor unit dysregulation following 15 days of unilateral lower limb immobilisation

Disuse atrophy, caused by situations of unloading such as limb immobilisation, causes a rapid yet diverging reduction in skeletal muscle function compared to muscle mass. While mechanistic insight into the loss of mass is well studied, deterioration of muscle function with a focus towards the neural input to muscle remains underexplored. This study aimed to determine the role of motor unit adaptation in disuse-induced neuromuscular deficits. Ten young, healthy male volunteers underwent 15 days of unilateral lower limb immobilisation with intramuscular EMG (iEMG) recorded from the vastus lateralis during knee extensor contractions normalised to maximal voluntary contraction (MVC) pre and post disuse. Muscle cross-sectional area was determined by ultrasound. Individual MUs were sampled and analysed for changes in MU discharge and MU potential (MUP) characteristics. Vastus lateralis (VL) CSA was reduced by approximately 15% which was exceeded by a two-fold decrease of 31% in muscle strength in the immobilised limb, with no change in either parameter in the non-immobilised VL. Parameters of MUP size were largely reduced with immobilisation, while neuromuscular junction (NMJ) transmission instability increased, and MU firing rate decreased at several contraction levels. All adaptations were observed in the immobilised limb only. These findings highlight impaired neural input following immobilisation reflected by suppressed MU discharge rate and instability of transmission at the NMJ which may underpin the disproportionate reductions of strength relative to muscle size. Key pointsO_LIMuscle mass and function decline rapidly in situations of disuse such as bed rest and limb immobilisation. C_LIO_LIThe reduction in muscle function commonly exceeds that of muscle mass, which may be associated with the dysregulation of neural input to the muscle. C_LIO_LIWe have used intramuscular electromyography to sample individual motor unit and near fibre potentials from the vastus lateralis following 15 days of unilateral limb immobilisation. Following disuse, the disproportionate loss of muscle strength when compared to size was associated with suppressed motor unit firing rate and increased markers of neuromuscular junction transmission instability. C_LIO_LIThese central and peripheral motor unit adaptations were observed at multiple contraction levels and in the immobilised limb only. Our findings demonstrate neural dysregulation as a key component of functional loss following muscle disuse in humans. C_LI

physiology↗

bioRxiv2022

Training induced improvements in knee extensor force accuracy are associated with reduced vastus lateralis motor unit firing variability

BackgroundMuscle force output during sustained submaximal isometric contractions fluctuates around an average value and is known to be influenced by variation in motor unit (MU) firing rates. MU firing rate variability seemingly reduces following exercise training interventions, however, much less is known with respect to peripheral MU properties. We therefore investigated whether targeted force accuracy training could lead to improved muscle functional capacity and control, in addition to determining any alterations of individual MU features. MethodsTen healthy participants (7 females, 3 males, 27{+/-}6 years, 170{+/-}8 cm, 69{+/-}16kg) underwent a 4-week supervised, unilateral, force accuracy training intervention. The coefficient of variation for force (FORCECoV) and sinusoidal wave force tracking accuracy (FORCESinu) were determined at 25% maximal voluntary contraction (MVC) pre- and post-training. Intramuscular electromyography was utilised to record individual MU potentials from the vastus lateralis (VL) muscles at 25% MVC during sustained contractions, pre- and post-training. ResultsKnee extensor muscle strength remained unchanged following training, with no improvements in unilateral leg-balance. FORCECoV and FORCESinu significantly improved in only the trained knee extensors by ~13% (p=0.01) and ~30% (p<0.0001) respectively. MU firing rate variability significantly reduced in the trained VL by ~16% (n=8; p=0.001), with no further alterations to MU firing rate or neuromuscular junction transmission instability. ConclusionOur results suggest muscle force control and tracking accuracy is a trainable characteristic in the knee extensors, which is likely explained by the reduction in MU firing rate variability apparent in the trained limb only.

physiology↗

bioRxiv2021

Motor unit recruitment strategies of the vastus lateralis according to sex

AimDespite men exhibiting greater muscle strength and fatigibility than women, it remains unclear if there are sex-based differences in muscle recruitment strategies e.g. motor unit (MU) recruitment and modulation of firing rate (FR) at normalised forces and during progressive increases in force. MethodsTwenty-nine healthy male and thirty-one healthy female participants (18-35 years) were studied. Intramuscular electromyography was used to record individual motor unit potentials (MUPs) and near fibre MUPs from the vastus lateralis (VL) during 10% and 25% maximum isometric voluntary contractions (MVC), and spike-triggered averaging was used to obtain motor unit number estimates (MUNE) of the VL. Multilevel mixed-effects linear regression models were used to investigate the effects of sex at each contraction level. ResultsMen exhibited greater muscle strength (p<0.001) and size (p<0.001) than women, with no difference in force steadiness at 10% or 25% MVC. Women had smaller MUs and higher FR at 10% MVC (both p<0.02), similar to that at 25% MVC in MU size (p=0.062) and FR (p=0.031). However, both sexes showed similar increases in MU size and FR when moving from low-to mid-level contractions. There were no sex differences in any near fibre MUP parameters or in MUNE. ConclusionIn the vastus lateralis, women produce muscle force via different neuromuscular recruitment strategies to men which is characterised by smaller MUs discharging at higher rates. However, similar strategies are employed to increase force production from low to moderate contractions. These findings of similar proportional increases between sexes support the use of mixed sex cohorts in studies of this nature. Key pointsO_LIIncreases in muscle force production are mediated by motor unit (MU) recruitment, and MU firing rate (FR). C_LIO_LIWomen are underrepresented in studies of human neuromuscular research and markedly differ to men in a number of aspects of neuromuscular function, yet little is known of the recruitment strategies of each. C_LIO_LIHere we demonstrate men and women have similar vastus lateralis MU number estimates, yet women recruit smaller MUs with higher FR than men at normalised contraction levels. However, increases in force are achieved via similar trajectories of MU recruitment and MU FR in men and women. C_LIO_LIAlthough men and women exhibit divergent neuromuscular recruitment strategies to achieve normalised forces, increases in force are achived similarly and support the inclusion of mixed sex cohorts in studies of this nature. C_LI

physiology↗

bioRxiv2020

Diet-Induced Vitamin D Deficiency Results in Reduced Skeletal Muscle Mitochondrial Respiration in C57BL/6J Mice

Vitamin D deficiency is known to be associated with symptoms of skeletal muscle myopathy including muscle weakness and fatigue. Recently, vitamin D related metabolites have been linked to the maintenance of mitochondrial function within skeletal muscle. However, current evidence is limited to in vitro models and the effects of diet-induced vitamin D deficiency upon skeletal muscle mitochondrial function in vivo have received little attention. In order to examine the role of vitamin D in the maintenance of mitochondrial function in vivo, we utilised an established model of diet-induced vitamin D deficiency in C57BL/6J mice. Mice were fed either a control (2,200 IU/kg) or a vitamin D deplete (0 IU/kg) diet for periods of 1-, 2- and 3-months. Skeletal muscle mitochondrial function and ADP sensitivity were assessed via high-resolution respirometry and mitochondrial protein content via immunoblotting. As a result of 3-month of diet-induced vitamin D deficiency, respiration supported via CI+IIP and ETC were 35% and 37% lower when compared to vitamin D replete mice (P < 0.05). Despite functional alterations, the protein expression of electron transfer chain subunits remained unchanged in response to dietary intervention (P > 0.05). In conclusion, we report that 3-months of diet-induced vitamin D deficiency reduced skeletal muscle mitochondrial function in C57BL/6J mice. Our data, when combined with previous in vitro observations, suggests that vitamin D mediated regulation of mitochondrial function may underlie the exacerbated muscle fatigue and performance deficits observed during vitamin D deficiency.

physiology↗

bioRxiv2019

The Vitamin D Receptor (VDR) Regulates Mitochondrial Function in C2C12 Myoblasts

Vitamin D deficiency has been linked to a reduction in skeletal muscle function and oxidative capacity, however, the mechanistic basis of these impairments are poorly understood. The biological actions of vitamin D are carried out via the binding of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) to the vitamin D receptor (VDR). Recent evidence has linked 1,25(OH)2D3 to the regulation of skeletal muscle mitochondrial function in vitro, however, little is known with regard to the role of the VDR in this process. To examine the regulatory role of the VDR in skeletal muscle mitochondrial function, we utilised lentiviral mediated shRNA silencing of the VDR in C2C12 myoblasts (VDR-KD) and examined mitochondrial respiration and protein content compared to shRNA scrambled control. VDR protein content was reduced by ~95% in myoblasts and myotubes (P < 0.001). VDR-KD myoblasts displayed a 30%, 30% and 36% reduction in basal, coupled and maximal respiration respectively (P < 0.05). This phenotype was maintained in VDR-KD myotubes, displaying a 34%, 33% and 48% reduction in basal, coupled and maximal respiration (P < 0.05). Furthermore, ATP production derived from oxidative phosphorylation (ATPox) was reduced by 20% suggesting intrinsic impairments within the mitochondria following VDR-KD. However, despite the observed functional decrements, mitochondrial protein content as well as markers of fusion and fission were unchanged. In summary, we highlight a direct role for the VDR in regulating skeletal muscle mitochondrial respiration in vitro, providing a potential mechanism as to how vitamin D deficiency might impact upon skeletal muscle oxidative capacity.

physiology↗