Skeletal muscle hypertrophy rewires glucose metabolism: An experimental investigation and systematic review.

BACKGROUND: Proliferating cancer cells shift their metabolism towards glycolysis, even in the presence of oxygen, to especially generate glycolytic intermediates as substrates for anabolic reactions. We hypothesize that a similar metabolic remodelling occurs during skeletal muscle hypertrophy. METHODS: We used mass spectrometry in hypertrophying C2C12 myotubes in vitro and plantaris mouse muscle in vivo and assessed metabolomic changes and the incorporation of the [U-13C6]glucose tracer. We performed enzyme inhibition of the key serine synthesis pathway enzyme phosphoglycerate dehydrogenase (Phgdh) for further mechanistic analysis and conducted a systematic review to align any changes in metabolomics during muscle growth with published findings. Finally, the UK Biobank was used to link the findings to population level. RESULTS: The metabolomics analysis in myotubes revealed insulin-like growth factor-1 (IGF-1)-induced altered metabolite concentrations in anabolic pathways such as pentose phosphate (ribose-5-phosphate/ribulose-5-phosphate: +40%; P = 0.01) and serine synthesis pathway (serine: -36.8%; P = 0.009). Like the hypertrophy stimulation with IGF-1 in myotubes in vitro, the concentration of the dipeptide l-carnosine was decreased by 26.6% (P = 0.001) during skeletal muscle growth in vivo. However, phosphorylated sugar (glucose-6-phosphate, fructose-6-phosphate or glucose-1-phosphate) decreased by 32.2% (P = 0.004) in the overloaded muscle in vivo while increasing in the IGF-1-stimulated myotubes in vitro. The systematic review revealed that 10 metabolites linked to muscle hypertrophy were directly associated with glycolysis and its interconnected anabolic pathways. We demonstrated that labelled carbon from [U-13C6]glucose is increasingly incorporated by ~13% (P = 0.001) into the non-essential amino acids in hypertrophying myotubes, which is accompanied by an increased depletion of media serine (P = 0.006). The inhibition of Phgdh suppressed muscle protein synthesis in growing myotubes by 58.1% (P < 0.001), highlighting the importance of the serine synthesis pathway for maintaining muscle size. Utilizing data from the UK Biobank (n = 450 243), we then discerned genetic variations linked to the serine synthesis pathway (PHGDH and PSPH) and to its downstream enzyme (SHMT1), revealing their association with appendicular lean mass in humans (P < 5.0e-8). CONCLUSIONS: Understanding the mechanisms that regulate skeletal muscle mass will help in developing effective treatments for muscle weakness. Our results provide evidence for the metabolic rewiring of glycolytic intermediates into anabolic pathways during muscle growth, such as in serine synthesis.

Effects of Diet Macronutrient Composition on Weight Loss during Caloric Restriction and Subsequent Weight Regain during Refeeding in Aging Mice.

Caloric restriction (CR) induces weight loss, but is associated with rapid weight regain upon return to ad libitum feeding. Our aim was to investigate effects of the macronutrient composition of the diet on weight loss and regain in elderly mice. Males, 18 months old, of the C57BL/6J strain were subjected to 4-week 30% CR followed by 4 weeks of ad libitum refeeding on either high-carb (HC), high-fat (HF) or high-protein (HP) diets (n = 22 each). Mice (n = 11) fed a chow diet ad libitum served as a control group (CON). Body mass and food intake were monitored daily. Twenty-four-hour indirect calorimetry was used to assess energy expenditure and substrate oxidation. Muscle and fat mass were evaluated with dissection of the tissues. Serum leptin and ghrelin levels were also measured. CR-induced weight loss did not differ between the diets. Weight regain was particularly fast for HF as mice overshot their initial weight by 12.8 ± 5.7% after 4-week refeeding when HC and HP mice reached the weight of the CON group. Weight regain strongly correlated with energy intake across the groups. The respiratory exchange ratio was lower in HF mice (0.81 ± 0.03) compared to HC (0.94 ± 0.06, p < 0.001), HP (0.89 ± 0.04, p < 0.001) and CON mice (0.91 ± 0.06, p < 0.01) during the refeeding. Serum leptin levels were higher in HF mice (1.03 ± 0.50 ng/mL) compared to HC (0.46 ± 0.14, p < 0.001), HP (0.63 ± 0.28, p < 0.05) or CON mice (0.41 ± 0.14, p < 0.001). Thus, CR induces similar weight loss in aging mice irrespective of the diet's macronutrient composition. An HF diet leads to excessive energy intake and pronounced gain in body fat in spite of increased fat oxidation and serum leptin during the refeeding after CR.

Stanniocalcin-2 inhibits skeletal muscle growth and is upregulated in functional overload-induced hypertrophy.

AIMS: Stanniocalcin-2 (STC2) has recently been implicated in human muscle mass variability by genetic analysis. Biochemically, STC2 inhibits the proteolytic activity of the metalloproteinase PAPP-A, which promotes muscle growth by upregulating the insulin-like growth factor (IGF) axis. The aim was to examine if STC2 affects skeletal muscle mass and to assess how the IGF axis mediates muscle hypertrophy induced by functional overload. METHODS: We compared muscle mass and muscle fiber morphology between Stc2-/- (n = 21) and wild-type (n = 15) mice. We then quantified IGF1, IGF2, IGF binding proteins -4 and -5 (IGFBP-4, IGFBP-5), PAPP-A and STC2 in plantaris muscles of wild-type mice subjected to 4-week unilateral overload (n = 14). RESULTS: Stc2-/- mice showed up to 10% larger muscle mass compared with wild-type mice. This increase was mediated by greater cross-sectional area of muscle fibers. Overload increased plantaris mass and components of the IGF axis, including quantities of IGF1 (by 2.41-fold, p = 0.0117), IGF2 (1.70-fold, p = 0.0461), IGFBP-4 (1.48-fold, p = 0.0268), PAPP-A (1.30-fold, p = 0.0154) and STC2 (1.28-fold, p = 0.019). CONCLUSION: Here we provide evidence that STC2 is an inhibitor of muscle growth upregulated, along with other components of the IGF axis, during overload-induced muscle hypertrophy.

Caloric Restriction per se Rather Than Dietary Macronutrient Distribution Plays a Primary Role in Metabolic Health and Body Composition Improvements in Obese Mice.

Caloric restriction (CR) is of key importance in combating obesity and its associated diseases. We aimed to examine effects of dietary macronutrient distribution on weight loss and metabolic health in obese mice exposed to CR. Male C57BL/6J mice underwent diet-induced obesity for 18 weeks. Thereafter mice were exposed to a 6-week CR for up to 40% on either low-fat diet (LFD; 20, 60, 20% kcal from protein, carbohydrate, fat), low-carb diet (LCD; 20, 20, 60% kcal, respectively) or high-pro diet (HPD; 35, 35, 30% kcal, respectively) (n = 16 each). Ten mice on the obesogenic diet served as age-matched controls. Body composition was evaluated by tissue dissections. Glucose tolerance, bloods lipids and energy metabolism were measured. CR-induced weight loss was similar for LFD and LCD while HPD was associated with a greater weight loss than LCD. The diet groups did not differ from obese controls in hindlimb muscle mass, but showed a substantial decrease in body fat without differences between them. Glucose tolerance and blood total cholesterol were weight-loss dependent and mostly improved in LFD and HPD groups during CR. Blood triacylglycerol was lowered only in LCD group compared to obese controls. Thus, CR rather than macronutrient distribution in the diet plays the major role for improvements in body composition and glucose control in obese mice. Low-carbohydrate-high-fat diet more successfully reduces triacylglycerol but not cholesterol levels compared to isocaloric high-carbohydrate-low-fat weight loss diets.

Effects of fasting on skeletal muscles and body fat of adult and old C57BL/6J mice.

Fasting improves metabolic health, but is also associated with loss of lean body mass. We investigated if old mice are less resistant to fasting-induce muscle wasting than adult mice. We compared changes in skeletal muscles and fat distribution in C57BL/6J mice subjected to 48-hour fasting at adult (6-month old) or old (24-month old) age. Old mice lost less weight (11.9 ± 1.5 vs 16.9 ± 2.8%, p < 0.001) and showed less (p < 0.01) pronounced muscle wasting than adult mice. Extensor digitorum longus (EDL) muscle force decreased only in adult mice after fasting. Serum IGF-1 levels were higher (p < 0.01) and showed greater (p < 0.01) decline in adult mice compared to old mice. Phosphorylation of 4EBP1 was reduced in the gastrocnemius muscles of adult mice only. Energy expenditure was slower in old mice and showed smaller fasting-induced decline than in adult mice when adjusted for variations in physical activity. There was a loss of fat mass in both age groups, but it was more pronounced in adult mice than old mice. Our results suggest that ageing-related decrease in metabolic rate protects old mice from skeletal muscle wasting during fasting.

Effects of ten-week 30% caloric restriction on metabolic health and skeletal muscles of adult and old C57BL/6J mice.

Caloric restriction (CR) can improve health, but the benefits are age-dependant. We studied effects of ten-week 30 % CR on skeletal muscles of adult (7-month old) and old (24-month old) C57BL/6 J mice. Old mice were heavier than adult mice (36.1 ± 4.0 g versus 32.9 ± 2.3 g, p < 0.05, respectively), but lost more weight (34.7 ± 6.0 % versus 23.9 ± 3.3 %, p < 0.001, respectively) during CR. Old mice did not differ from adult mice in extent of hind-limb muscle wasting or improvement in glucose tolerance after CR. Ageing and CR had an additive effect on increase in percentage of type 1 fibres in the soleus (SOL) muscle. CR was associated with greater atrophy of fast-twitch extensor digitorum longus (EDL) compared to slow-twitch SOL muscle. Old mice showed reduced gene expression of lysosomal markers, p62 and LC3B, while CR tended to upregulate the proteolysis genes. CR was also associated with increase in specific force of EDL muscle, but did not affect it in SOL muscle. In summary, ten-week CR induces only limited improvements in skeletal muscle function, but leads to significant muscle wasting and weakness in both adult and old mice.

Hypocaloric Low-Carbohydrate and Low-Fat Diets with Fixed Protein Lead to Similar Health Outcomes in Obese Mice.

OBJECTIVE: It is controversial whether low-carbohydrate diets are better suited for weight control and metabolic health than high-carbohydrate diets. This study examined whether these diets induce different improvements in body composition and glucose tolerance in obese mice during caloric restriction (CR). METHODS: Male C57BL/6J mice were fed an obesogenic diet ad libitum for 18 weeks and then subjected to 6-week progressive CR of up to 40%, using either a low-fat or low-carbohydrate diet with equal protein content. Mice fed a regular chow diet ad libitum served as controls. Body mass, hindlimb muscle mass, fat mass, energy expenditure, and glucose tolerance were compared between the groups. RESULTS: Initially low-fat and low-carbohydrate groups had similar body mass, which was 30% greater compared with controls. CR induced similar weight loss in low-fat and low-carbohydrate groups. This weight loss was mainly due to fat loss in both groups. Energy expenditure of freely moving mice did not differ between the groups. Glucose tolerance improved compared with the values before CR and in controls but did not differ between the diets. CONCLUSIONS: Dietary carbohydrate or fat content does not affect improvements in body composition and metabolic health in obese mice exposed to CR with fixed energy and protein intake.

Effect of fasting on body composition and proteolysis gene expression in skeletal muscles and liver of BEH+/+ and BEL mice.

Fasting improves health, but can cause muscle weakness. We assessed body composition in 21-week old males of Berlin high (BEH+/+) and Berlin low (BEL) strains after two bouts of 48-h or 40-h of fasting with 5-day refeeding in between, respectively. BEH+/+ mice tended to loose less weight than BEL in bout 1 and 2 (16.0 ± 2.7 versus 23.5 ± 2.9%, p < 0.001 and 17.1 ± 3.4 versus 20.4 ± 3.4%, p = 0.17, respectively). In spite of greater serum IGF-1 and body fat levels, BEH+/+ mice showed more severe muscle atrophy, but less marked liver wasting and fat depletion than BEL mice. BEH+/+ mice also showed smaller increases in expression of p62, Atrogin-1, and Mstn genes in skeletal muscles. In summary, BEL mice show resistance to fasting-induced muscle wasting in spite of low serum IGF-1 levels and high expression of genes associated with muscle atrophy.

Myostatin dysfunction is associated with lower physical activity and reduced improvements in glucose tolerance in response to caloric restriction in Berlin high mice.

Myostatin is an inhibitor of skeletal muscle growth and might be involved in adaptations to caloric restriction (CR). We compared responses to 12-week 30% CR in male mice of Berlin high strain with myostatin dysfunction (BEH) and wild-type myostatin (BEH+/+). BEH mice were heavier than BEH+/+ mice (58.8 ±â€¯2.0 versus 53.1 ±â€¯2.7 g, p < 0.001), had 1.8-fold greater hind limb muscle mass and were less (p < 0.05) physically active when fed ad libitum. After CR, BEH and BEH+/+ strains experienced similar weight loss (24.7 ±â€¯5.7 versus 20.6 ±â€¯6.5%, p > 0.05, respectively) and decreases (p < 0.001) in plasma IGF-1 and total cholesterol, but loss of hind limb muscle mass was greater (p < 0.001) in BEH mice than BEH+/+ mice. BEH mice had better (p < 0.001) glucose tolerance and showed smaller (p < 0.05) improvements of it than BEH+/+ mice after CR (1038.2 ±â€¯174.7 versus 744.4 ±â€¯95.8 glucose mM× 120 min, p < 0.01 for BEH; 1365.8 ±â€¯218.5 versus 831.5 ±â€¯134.4 glucose mM ×120 min, p < 0.001, for BEH+/+, respectively). In summary, myostatin dysfunction is associated with muscle hypertrophy and high glucose tolerance, but greater muscle wasting and smaller improvements in glucose tolerance in response to CR.

Myostatin dysfunction does not protect from fasting-induced loss of muscle mass in mice.

OBJECTIVES: The aim of the study was to investigate if myostatin dysfunction can ameliorate fasting-induced muscle wasting. METHODS: 18-week old males from Berlin high (BEH) strain with myostatin dysfunction and wild type myostatin (BEH+/+) strain were subjected to 48-h food deprivation (FD). Changes in body composition as well as contractile properties of soleus (SOL) and extensor digitorum longus (EDL) muscles were studied. RESULTS: BEH mice were heavier than BEH+/+ mice (56.0±2.5 vs. 49.9±2.8 g, P<0.001, respectively). FD induced similar loss of body mass in BEH and BEH+/+ mice (16.6±2.4 vs. 17.4±2.2%, P>0.05), but only BEH mice experienced wasting of the gastrocnemius, tibialis anterior and plantaris muscles. FD induced a marked decrease in specific muscle force of SOL. EDL of BEH mice tended to be protected from this decline. CONCLUSION: Myostatin dysfunction does not protect from loss of muscle mass during fasting.

Corrigendum to "Low Citrate Synthase Activity Is Associated with Glucose Intolerance and Lipotoxicity".

[This corrects the article DOI: 10.1155/2019/8594825.].

Low Citrate Synthase Activity Is Associated with Glucose Intolerance and Lipotoxicity.

Citrate synthase (CS) is a key mitochondrial enzyme. The aim of this study was to test the hypothesis that low CS activity impairs the metabolic health of mice fed a high fat diet (HFD) and promotes palmitate-induced lipotoxicity in muscle cells. C57BL/6J (B6) mice and congenic B6.A-(rs3676616-D10Utsw1)/KjnB6 (B6.A), a strain which carries the A/J allele of CS on the B6 strain background, were fed HFD (45% kcal from fat) for 12 weeks. C2C12 mouse muscle cells were used to investigate effects of CS knockdown on cell viability and signalling after incubation in 0.8 mM palmitate. CS activity, but not that of ß-hydroxyacyl-coenzyme-A dehydrogenase was lower in the gastrocnemius muscle and heart of B6.A mice compared to B6 mice (P < 0.001). During HFD feeding, glucose tolerance of mice decreased progressively and to a greater extent in B6.A females compared to B6 females, with males showing a similar trend. Body weight and fat gain did not differ between B6.A and B6 mice. After an 18 h incubation in 0.8 mM palmitate C2C12 muscle cells with ∼50% shRNA mediated reduction in CS activity showed lower (P < 0.001) viability and increased (P < 0.001) levels of cleaved caspase-3 compared to the scramble shRNA treated C2C12 cells. A/J strain variant of CS is associated with low enzyme activity and impaired metabolic health. This could be due to impaired lipid metabolism in muscle cells.

Pre-Exercise Rehydration Attenuates Central Fatigability during 2-Min Maximum Voluntary Contraction in Hyperthermia.

Background and objectives: Hyperthermia with dehydration alters several brain structure volumes, mainly by changing plasma osmolality, thus strongly affecting neural functions (cognitive and motor). Here, we aimed to examine whether the prevention of significant dehydration caused by passively induced whole-body hyperthermia attenuates peripheral and/or central fatigability during a sustained 2-min isometric maximal voluntary contraction (MVC). Materials and Methods: Ten healthy and physically active adult men (21 ± 1 years of age) performed an isometric MVC of the knee extensors for 2 min (2-min MVC) under control (CON) conditions, after passive lower-body heating that induced severe whole-body hyperthermia (HT, Tre > 39 °C) with dehydration (HT-D) and after HT with rehydration (HT-RH). Results: In the HT-D trial, the subjects lost 0.94 ± 0.15 kg (1.33% ± 0.13%) of their body weight; in the HT-RH trial, their body weight increased by 0.1 ± 0.42 kg (0.1% ± 0.58%). After lower-body heating, the HT-RH trial (vs. HT-D trial) was accompanied by a significantly lower physiological stress index (6.77 ± 0.98 vs. 7.40 ± 1.46, respectively), heart rate (47.8 ± 9.8 vs. 60.8 ± 13.2 b min-1, respectively), and systolic blood pressure (-12.52 ± 5.1 vs. +2.3 ± 6.4, respectively). During 2-min MVC, hyperthermia (HT-D; HT-RH) resulted in greater central fatigability compared with the CON trial. The voluntary activation of exercising muscles was less depressed in the HT-RH trial compared with the HT-D trial. Over the exercise period, electrically (involuntary) induced torque decreased less in the HT-D trial than in the CON and HT-RH trials. Conclusions: Our results suggest that pre-exercise rehydration might have the immediate positive effect of reducing physiological thermal strain, thus attenuating central fatigability even when exercise is performed during severe (Tre > 39 °C) HT, induced by passive warming of the lower body.

Effects of myostatin on the mechanical properties of muscles during repeated active lengthening in the mouse.

The aim of the present study was to investigate how myostatin dysfunction affects fast and slow muscle stiffness and viscosity during severe repeated loading. Isolated extensor digitorum longus (EDL) and soleus muscles of young adult female mice of the BEH (dysfunctional myostatin) and BEH+/+ (functional myostatin) strains were subjected to 100 contraction-stretching loading cycles during which contractile and mechanical properties were assessed. BEH mice exhibited greater exercise-induced muscle damage, although the effect was muscle- and age-dependent and limited to the early phases of simulated exercise. The relative reduction of the EDL muscle isometric force recorded during the initial 10-30 loading cycles was greater in BEH mice than in BEH+/+ mice and exceeded that of the soleus muscle of either strain. The induced damage was associated with lower muscle stiffness. The effects of myostatin on the mechanical properties of muscles depend on muscle type and maturity.

H55N polymorphism is associated with low citrate synthase activity which regulates lipid metabolism in mouse muscle cells.

The H55N polymorphism in the Cs gene of A/J mice has been linked to low activity of the enzyme in skeletal muscles. The aim of the study was to test this hypothesis and examine effects of low citrate synthase (CS) activity on palmitate metabolism in muscle cells. Results of the study showed that carriers of the wild type (WT) Cs (C57BL/6J and Balb/cByJ mouse strains) had higher CS activity (p < 0.01) than carriers of the A/J variant (B6.A-(rs3676616-D10Utsw1)/KjnB6 and A/J mouse strains) in the heart, liver and gastrocnemius muscle. Furthermore, the recombinant CS protein of WT showed higher CS activity than the A/J variant. In C2C12 muscle cells the shRNA mediated 47% knockdown of CS activity reduced the rate of fatty acid oxidation compared to the control cells. In summary, our results are consistent with the hypothesis that H55N substitution causes a reduction in CS activity. Furthermore, low CS activity interferes with metabolic flexibility of muscle cells.

Age-related changes in the effects of strength training on lower leg muscles in healthy individuals measured using MRI.

BACKGROUND: We previously measured the rate of regaining muscle strength during rehabilitation of lower leg muscles in patients following lower leg casting. Our primary aim in this study was to measure the rate of gain of strength in healthy individuals undergoing a similar training regime. Our secondary aim was to test the ability of MRI to provide a biomarker for muscle function. METHODS: Men and women were recruited in three age groups: 20-30, 50-65 and over 70 years. Their response to resistance training of the right lower leg twice a week for 8 weeks was monitored using a dynamometer and MRI of tibialis anterior, soleus and gastrocnemius muscles at 2 weekly intervals to measure muscle size (anatomical cross-sectional area (ACSA)) and quality (T2 relaxation). Forty-four volunteers completed the study. RESULTS: Baseline strength declined with age. Training had no effect in middle-aged females or in elderly men in dorsiflexion. Other groups significantly increased both plantarflexion and dorsiflexion strength at rates up to 5.5 N m week-1 in young females in plantarflexion and 1.25 N m week-1 in young males in dorsiflexion. No changes were observed in ACSA or T2 in any age group in any muscle. CONCLUSION: Exercise training improves muscle strength in males at all ages except the elderly in dorsiflexion. Responses in females were less clear with variation across age and muscle groups. These results were not reflected in simple MRI measures that do not, therefore, provide a good biomarker for muscle atrophy or the efficacy of rehabilitation.

Forced Running Endurance Is Influenced by Gene(s) on Mouse Chromosome 10.

Phenotypic diversity between laboratory mouse strains provides a model for studying the underlying genetic mechanisms. The A/J strain performs poorly in various endurance exercise models. The aim of the study was to test if endurance capacity and contractility of the fast- and slow-twitch muscles are affected by the genes on mouse chromosome 10. The C57BL/6J (B6) strain and C57BL/6J-Chr 10A/J/NaJ (B6.A10) consomic strain which carries the A/J chromosome 10 on a B6 strain background were compared. The B6.A10 mice compared to B6 were larger in body weight (p < 0.02): 27.2 ± 1.9 vs. 23.8 ± 2.7 and 23.4 ± 1.9 vs. 22.9 ± 2.3 g, for males and females, respectively, and in male soleus weight (p < 0.02): 9.7 ± 0.4 vs. 8.6 ± 0.9 mg. In the forced running test the B6.A10 mice completed only 64% of the B6 covered distance (p < 0.0001). However, there was no difference in voluntary wheel running (p = 0.6) or in fatigability of isolated soleus (p = 0.24) or extensor digitorum longus (EDL, p = 0.7) muscles. We conclude that chromosome 10 of the A/J strain contributes to reduced endurance performance. We also discuss physiological mechanisms and methodological aspects relevant to interpretation of these findings.

Myostatin dysfunction impairs force generation in extensor digitorum longus muscle and increases exercise-induced protein efflux from extensor digitorum longus and soleus muscles.

Myostatin dysfunction promotes muscle hypertrophy, which can complicate assessment of muscle properties. We examined force generating capacity and creatine kinase (CK) efflux from skeletal muscles of young mice before they reach adult body and muscle size. Isolated soleus (SOL) and extensor digitorum longus (EDL) muscles of Berlin high (BEH) mice with dysfunctional myostatin, i.e., homozygous for inactivating myostatin mutation, and with a wild-type myostatin (BEH+/+) were studied. The muscles of BEH mice showed faster (P < 0.01) twitch and tetanus contraction times compared with BEH+/+ mice, but only EDL displayed lower (P < 0.05) specific force. SOL and EDL of age-matched but not younger BEH mice showed greater exercise-induced CK efflux compared with BEH+/+ mice. In summary, myostatin dysfunction leads to impairment in muscle force generating capacity in EDL and increases susceptibility of SOL and EDL to protein loss after exercise.

Efflux of creatine kinase from isolated soleus muscle depends on age, sex and type of exercise in mice.

Elevated plasma creatine kinase (CK) activity is often used as an indicator of exercise-induced muscle damage. Our aim was to study effects of contraction type, sex and age on CK efflux from isolated skeletal muscles of mice. The soleus muscle (SOL) of adult (7.5-month old) female C57BL/6J mice was subjected to either 100 passive stretches, isometric contractions or eccentric contractions, and muscle CK efflux was assessed after two-hour incubation in vitro. SOL of young (3-month old) male and female mice was studied after 100 eccentric contractions. For adult females, muscle CK efflux was larger (p < 0.05) after eccentric contractions than after incubation without exercise (698 ± 344 vs. 268 ± 184 mU·h(-1), respectively), but smaller (p < 0.05) than for young females after the same type of exercise (1069 ± 341 mU·h(-1)). Eccentric exercise-induced CK efflux was larger in muscles of young males compared to young females (2046 ± 317 vs 1069 ± 341 mU · h(-1), respectively, p < 0.001). Our results show that eccentric contractions induce a significant increase in muscle CK efflux immediately after exercise. Isolated muscle resistance to exercise-induced CK efflux depends on age and sex of mice. Key pointsMuscle lengthening contractions induce the highest CK efflux in vitro compared with similar protocol of isometric contractions or passive stretches.Muscle CK efflux in vitro is applicable in studying changes of sarcolemma permeability/integrity, a proxy of muscle damage, in response to muscle contractile activity.Isolated muscle resistance to exercise-induced CK efflux is greater in female compared to male mice of young age and is further increased in adult female mice.

Regenerated soleus muscle shows reduced creatine kinase efflux after contractile activity in vitro.

Regenerated skeletal muscles show less muscle damage after strenuous muscle exercise. The aim of the studies was to investigate if the regeneration is associated with reduced muscle creatine kinase (CK) efflux immediately after the exercise. Cryolesion was applied to the soleus muscle of 3-month-old C57BL/6J male mice. Then total CK efflux was assessed in vitro in the regenerated muscles without exercise or after 100 eccentric contractions. The same measurements were performed in the control muscles, which were not exposed to cryolesion. Regenerated muscles generated weaker (P < 0.05) twitches, but stronger (P < 0.05) 150-Hz and 300-Hz tetani with prolonged (P < 0.01) contraction times compared with the control muscles. There was no difference between regenerated and control muscles in the total CK efflux without exercise, but only control muscles showed an increase (P < 0.001) in the CK efflux after the exercise. Our results suggest that muscle regeneration is associated with modulation of contractile properties and improvement in muscle resistance to damage after eccentric exercise.