Lars Grøndahl Hvid

Effects of aging on human skeletal muscle after immobilization and retraining

Inactivity is a recognized compounding factor in sarcopenia and muscle weakness in old age. However, while the negative effects of unloading on skeletal muscle in young individuals are well elucidated, only little is known about the consequence of immobilization and the regenerative capacity in elderly individuals. Thus the aim of this study was to examine the effect of aging on changes in muscle contractile properties, specific force, and muscle mass characteristics in 9 old (61-74 yr) and 11 young men (21-27 yr) after 2 wk of immobilization and 4 wk of retraining. Both young and old experienced decreases in maximal muscle strength, resting twitch peak torque and twitch rate of force development, quadriceps muscle volume, pennation angle, and specific force after 2 wk of immobilization (P < 0.05). The decline in quadriceps volume and pennation angle was smaller in old compared with young (P < 0.05). In contrast, only old men experienced a decrease in quadriceps activation. After retraining, both young and old regained their initial muscle strength, but old had smaller gains in quadriceps volume compared with young, and pennation angle increased in young only (P < 0.05). The present study is the first to demonstrate that aging alters the neuromuscular response to short-term disuse and recovery in humans. Notably, immobilization had a greater impact on neuronal motor function in old individuals, while young individuals were more affected at the muscle level. In addition, old individuals showed an attenuated response to retraining after immobilization compared with young individuals.

Proliferation of myogenic stem cells in human skeletal muscle in response to low-load resistance training with blood flow restriction

In the last decade muscle training performed using a combination of low external loads and partial restriction of blood flow to the exercising limb has gained increasing interest, since it leads to significant gains in muscle strength and muscle mass. The cellular mechanisms responsible for the muscular adaptations induced by this training paradigm are not fully understood. This study shows that 3 weeks of high‐frequency, low‐intensity muscle exercise with partial blood flow restriction induces increases in maximal muscle strength accompanied by highly marked gains in muscle fibre size. Furthermore, the results indicate that these muscular adaptations rely on a considerable upregulation in myogenic satellite cells number, resulting in nuclear addition to the exercised myofibres. The results contribute to a better understanding of the physiological mechanisms underlying the gain in muscle strength and muscle mass observed with blood flow restricted low‐intensity resistance exercise.

Effects of aging on muscle mechanical function and muscle fiber morphology during short-term immobilization and subsequent retraining

Very little attention has been given to the combined effects of aging and disuse as separate factors causing deterioration in muscle mechanical function. Thus the purpose of this study was to investigate the effects of 2 wk of immobilization followed by 4 wk of retraining on knee extensor muscle mechanical function (e.g., maximal strength and rapid force capacity) and muscle fiber morphology in 9 old (OM: 67.3 ± 1.3 yr) and 11 young healthy men (YM: 24.4 ± 0.5 yr) with comparable levels of physical activity. Following immobilization, OM demonstrated markedly larger decreases in rapid force capacity (i.e., rate of force development, impulse) than YM (∼ 20-37 vs. ∼ 13-16%; P < 0.05). In contrast, muscle fiber area decreased in YM for type I, IIA, and IIx fibers (∼ 15-30%; P < 0.05), whereas only type IIa area decreased in OM (13.2%; P < 0.05). Subsequent retraining fully restored muscle mechanical function and muscle fiber area in YM, whereas OM showed an attenuated recovery in muscle fiber area and rapid force capacity (tendency). Changes in maximal isometric and dynamic muscle strength were similar between OM and YM. In conclusion, the present data reveal that OM may be more susceptible to the deleterious effects of short-term muscle disuse on muscle fiber size and rapid force capacity than YM. Furthermore, OM seems to require longer time to recover and regain rapid muscle force capacity, which may lead to a larger risk of falling in aged individuals after periods of short-term disuse.

Aging affects the transcriptional regulation of human skeletal muscle disuse atrophy.

Important insights concerning the molecular basis of skeletal muscle disuse-atrophy and aging related muscle loss have been obtained in cell culture and animal models, but these regulatory signaling pathways have not previously been studied in aging human muscle. In the present study, muscle atrophy was induced by immobilization in healthy old and young individuals to study the time-course and transcriptional factors underlying human skeletal muscle atrophy. The results reveal that irrespectively of age, mRNA expression levels of MuRF-1 and Atrogin-1 increased in the very initial phase (2–4 days) of human disuse-muscle atrophy along with a marked reduction in PGC-1α and PGC-1β (1–4 days) and a ∼10% decrease in myofiber size (4 days). Further, an age-specific decrease in Akt and S6 phosphorylation was observed in young muscle within the first days (1–4 days) of immobilization. In contrast, Akt phosphorylation was unchanged in old muscle after 2 days and increased after 4 days of immobilization. Further, an age-specific down-regulation of MuRF-1 and Atrogin-1 expression levels was observed following 2 weeks of immobilization, along with a slowing atrophy response in aged skeletal muscle. Neither the immediate loss of muscle mass, nor the subsequent age-differentiated signaling responses could be explained by changes in inflammatory mediators, apoptosis markers or autophagy indicators. Collectively, these findings indicate that the time-course and regulation of human skeletal muscle atrophy is age dependent, leading to an attenuated loss in aging skeletal muscle when exposed to longer periods of immobility-induced disuse.

Ageing is associated with diminished muscle re-growth and myogenic precursor cell expansion early after immobility-induced atrophy in human skeletal muscle

•  Elderly individuals require a prolonged recovery phase in order to return to initial muscle mass levels following short‐term immobilisation. •  The cellular mechanisms responsible for the attenuated re‐growth and associated molecular signalling processes in ageing human skeletal muscle are not fully understood. •  The main study finding was the observation of a less marked muscle mass recovery after immobilisation in elderly compared to young individuals that was paralleled by an elevation in myogenic precursor cell content in young individuals only, whereas the elderly failed to demonstrate any change in myogenic precursor cells. •  No age‐related differences were observed in the expression of major myogenic regulating factors known to promote skeletal muscle hypertrophy or satellite cell proliferation (IGF‐1Ea, MGF, MyoD1, myogenin, HGF gene products). •  In contrast, the expression of myostatin demonstrated a more pronounced up‐regulation following immobilisation along with an attenuated down‐regulation in response to reloading in older compared to young individuals, which may have contributed to the present lack of satellite cell proliferation in ageing muscle.

Effects of ageing on single muscle fibre contractile function following short-term immobilisation

Non‐Technical Summary  The contractile function of human single muscle fibres is of particular importance for whole muscle contractile function. Yet, whereas ageing and short‐term disuse (immobilisation) separately have been shown to impair single fibre contractile function, very little attention has been given to their combined effects. We show that 2 weeks of lower limb immobilisation reduces force and specific force (force per cross‐sectional area) of both slow and fast single muscle fibres and that this occurred to a similar extent in young and old individuals. In contrast, disuse led to reduced Ca2+ sensitivity in fast fibres of young and in slow fibres of old, respectively. These results help us to better understand the underlying physiological mechanisms responsible for the deleterious effects of short‐term disuse on whole muscle contractile function in both young and old.

Aging impairs the recovery in mechanical muscle function following 4 days of disuse.

As aged individuals are frequently exposed to short-term disuse caused by disease or musculoskeletal injury, it is important to understand how short-term disuse and subsequent retraining affect lower limb mechanical muscle function. The purpose of the present study was, therefore, to investigate the effect of 4 days of lower limb disuse followed by 7 days of active recovery on mechanical muscle function of the knee extensors in young (24.3±0.9 years, n=11) and old (67.2±1.0 years, n=11) recreationally active healthy males. Slow and moderate dynamic muscle strength were assessed using isokinetic dynamometry (60 and 180° s(-1), respectively) along with isometric muscle strength and rapid muscle force capacity examined as contractile rate of force development (RFD), Impulse, and relative RFD (rRFD) during the initial phase of contraction (100 ms time interval relative to onset of contraction). Prior to disuse, marked age-related differences (p<0.05) were observed in isometric and dynamic muscle strength (~35%) as well as in RFD and Impulse (~39%). Following disuse, young and old individuals experienced comparable decrements (p<0.05) in isometric strength (~9%), slow dynamic strength (~13%), and RFD and Impulse (~19%), whereas old individuals only experienced decrements (p<0.05) in moderate dynamic strength (12%) and rRFD (~17%). Following recovery, all measures of mechanical muscle function were restored in young individuals compared to pre-disuse values, while isometric, slow and moderate dynamic muscle strength remained suppressed (p<0.05) in old individuals (~8%) along with a tendency to suppressed RFD100 (p=0.068). In conclusion, 4 days of lower limb disuse led to marked decrements in knee extensor mechanical muscle function in both young and old individuals, yet with greater decrements observed in moderate dynamic strength and rapid muscle force capacity in old individuals. While 7 days of recovery - including free ambulation, one test session and a single session of strength training - was sufficient to restore mechanical muscle function in young individuals, old individuals appeared to have an impaired ability to fully recover as evidenced by suppressed values of isometric and dynamic muscle strength and rapid muscle force capacity.

Subcellular localization-dependent decrements in skeletal muscle glycogen and mitochondria content following short-term disuse in young and old men

Previous studies have shown that skeletal muscle glycogen and mitochondria are distributed in distinct subcellular localizations, but the role and regulation of these subcellular localizations are unclear. In the present study, we used transmission electron microscopy to investigate the effect of disuse and aging on human skeletal muscle glycogen and mitochondria content in subsarcolemmal (SS), intermyofibrillar (IMF), and intramyofibrillar (intra) localizations. Five young (∼23 yr) and five old (∼66 yr) recreationally active men had their quadriceps muscle immobilized for 2 wk by whole leg casting. Biopsies were obtained from m. vastus lateralis before and after the immobilization period. Immobilization induced a decrement of intra glycogen content by 54% (P < 0.001) in both age groups and in two ultrastructurally distinct fiber types, whereas the content of IMF and SS glycogen remained unchanged. A localization-dependent decrease (P = 0.03) in mitochondria content following immobilization was found in both age groups, where SS mitochondria decreased by 33% (P = 0.02), superficial IMF mitochondria decreased by 20% (P = 0.05), and central IMF mitochondria remained unchanged. In conclusion, our findings demonstrate a localization-dependent adaptation to immobilization in glycogen and mitochondria content of skeletal muscles of both young and old individuals. Specifically, this suggests that short-term disuse preferentially affects glycogen particles located inside the myofibrils and that mitochondria volume plasticity can be dependent on the distance to the fiber border.

Four days of muscle disuse impairs single fiber contractile function in young and old healthy men.

The purpose of the study was to investigate the effects of 4 days of disuse (knee brace) on contractile function of isolated vastus lateralis fibers (n=486) from 11 young (24.3±0.9 yrs) and 11 old (67.2±1.0 yrs) healthy men having comparable levels of physical activity. Prior to disuse single fiber specific force (maximal Ca(2+)-activated force per cross-sectional area) was lower in MHC I vs. IIa fibers (p<0.05) both in young (44%) and old (32%), and specific force of MHC IIa fibers was lower in old vs. young (19%, p<0.05). Further, Ca(2+) sensitivity was higher in MHC I vs. IIa fibers (p<0.05) in both age groups. Following disuse single fiber specific force decreased (p<0.05) in MHC I fibers in young (-19%) and old (-17%), and in MHC IIa fibers in young (-21%) but not in old (-11%; ns). The extent of these decreases did not differ between young and old. Ca(2+) sensitivity decreased (p<0.05) in MHC I fibers in old (-0.11 pCa units) but not in young (-0.08 pCa units; ns). The extent of these decreases was greater in old vs. young (p<0.05). In conclusion, 4 days of lower limb disuse led to marked impairments in single muscle fiber specific force independently of age, while changes in Ca(2+) sensitivity were dependent on age and MHC isoform composition. The present findings stress the importance of determining and implementing effective preventive and rehabilitative approaches for old individuals exposed to as little as 4 days of disuse.

Muscle Glycogen Content Modifies SR Ca2 + Release Rate in Elite Endurance Athletes.

PURPOSE The aim of the present study was to investigate the influence of muscle glycogen content on sarcoplasmic reticulum (SR) function and peak power output (Wpeak) in elite endurance athletes. METHODS Fourteen highly trained male triathletes (VO2max = 66.5 ± 1.3 mL O2·kg·min), performed 4 h of glycogen-depleting cycling exercise (HRmean = 73% ± 1% of maximum). During the first 4 h of recovery, athletes received either water (H2O) or carbohydrate (CHO), separating alterations in muscle glycogen content from acute changes affecting SR function and performance. Thereafter, all subjects received CHO-enriched food for the remaining 20-h recovery period. RESULTS Immediately after exercise, muscle glycogen content and SR Ca release rate was reduced to 32% ± 4% (225 ± 28 mmol·kg dw) and 86% ± 2% of initial levels, respectively (P < 0.01). Glycogen markedly recovered after 4 h of recovery with CHO (61% ± 2% of preexercise) and SR Ca release rate returned to preexercise level. However, in the absence of CHO during the first 4 h of recovery, glycogen and SR Ca release rate remained depressed, with the normalization of both parameters at the end of the 24 h of recovery after receiving a CHO-enriched diet. Linear regression demonstrated a significant correlation between SR Ca release rate and muscle glycogen content (P < 0.01, r = 0.30). The 4 h of cycling exercise reduced Wpeak by 5.5%-8.9% at different cadences (P < 0.05), and Wpeak was normalized after 4 h of recovery with CHO, whereas Wpeak remained depressed (P < 0.05) after water provision. Wpeak was fully recovered after 24 h in both the H2O and the CHO group. CONCLUSION In conclusion, the present results suggest that low muscle glycogen depresses muscle SR Ca release rate, which may contribute to fatigue and delayed recovery of Wpeak 4 h postexercise.