<scp>l</scp>‐glutamine Improves Skeletal Muscle Cell Differentiation and Prevents Myotube Atrophy After Cytokine (TNF‐α) Stress Via Reduced p38 MAPK Signal Transduction

Girven, Matthew; Dugdale, Hannah F.; Owens, Daniel J.; Hughes, David C.; Stewart, Claire E.; Sharples, Adam P.

doi:10.1002/jcp.25380

Cited by 44 publications

(35 citation statements)

References 86 publications

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“…Cells were then seeded at 90,000 cells mL –1 in 2 mL of growth media [GM; 4.5 g L –1 glucose DMEM, which included 4 m m l ‐glutamine, 10% hiFBS, 10% hiNBCS, supplemented with an additional 2 m m l ‐glutamine (Girven et al . ), 100 U mL –1 penicillin, 100 µg mL –1 streptomycin, 0.5 mg mL –1 6‐aminocaproic acid, 50 µ m l ‐ascorbic acid, 50 µ m l ‐proline] onto each sylgard coated (Sylgard 184 Elastomer Kit; Dow Corning, Midland, MI, USA) 35 mm culture dish containing a pre‐polymerized (10 min at room temperature and 1 h at 37°C) fibrin gel. The fibrin gel was made up of 500 µL of thrombin‐solution (10 U mL –1 thrombin, 8 µL mL –1 aprotinin at 10 mg mL –1 ) and 200 µL of fibrinogen at 20 mg mL –1 ).…”

Section: Methodsmentioning

confidence: 99%

“…Briefly, C2C12 cells were grown on T75 flasks to ß80% confluence. Cells were then seeded at 90,000 cells mL -1 in 2 mL of growth media [GM; 4.5 g L -1 glucose DMEM, which included 4 mM L-glutamine, 10% hiFBS, 10% hiNBCS, supplemented with an additional 2 mM L-glutamine (Girven et al 2016), 100 U mL -1 penicillin, 100 µg mL -1 streptomycin, 0.5 mg mL -1 6-aminocaproic acid, 50 µM L-ascorbic acid, 50 µM L-proline] onto each sylgard coated ( Figure 3. UBR5 gene expression and protein abundance after skeletal muscle atrophy and recovery following nerve crush injury A, muscle weight (mg) of the gastrocnemius/GSTC in mice after nerve crush injury (NCI) over a period of atrophy (3-14 days) and recovery (14-60 days) of muscle mass.…”

Section: Mechanical Loading Of Bioengineered Skeletal Muscle In Vitromentioning

confidence: 99%

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UBR5 is a novel E3 ubiquitin ligase involved in skeletal muscle hypertrophy and recovery from atrophy

Seaborne

Hughes

Turner

et al. 2019

The Journal of Physiology

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Key points We have recently identified that a HECT domain E3 ubiquitin ligase, named UBR5, is altered epigenetically (via DNA methylation) after human skeletal muscle hypertrophy, where its gene expression is positively correlated with increasing lean leg mass after training and retraining. In the present study we extensively investigate this novel and uncharacterised E3 ubiquitin ligase (UBR5) in skeletal muscle atrophy, recovery from atrophy and injury, anabolism and hypertrophy. We demonstrated that UBR5 was epigenetically altered via DNA methylation during recovery from atrophy. We also determined that UBR5 was alternatively regulated versus well characterised E3 ligases, MuRF1/MAFbx, at the gene expression level during atrophy, recovery from atrophy and hypertrophy. UBR5 also increased at the protein level during recovery from atrophy and injury, hypertrophy and during human muscle cell differentiation. Finally, in humans, genetic variations of the UBR5 gene were strongly associated with larger fast‐twitch muscle fibres and strength/power performance versus endurance/untrained phenotypes. Abstract We aimed to investigate a novel and uncharacterized E3 ubiquitin ligase in skeletal muscle atrophy, recovery from atrophy/injury, anabolism and hypertrophy. We demonstrated an alternate gene expression profile for UBR5 vs. well characterized E3‐ligases, MuRF1/MAFbx, where, after atrophy evoked by continuous‐low‐frequency electrical‐stimulation in rats, MuRF1/MAFbx were both elevated, yet UBR5 was unchanged. Furthermore, after recovery of muscle mass post TTX‐induced atrophy in rats, UBR5 was hypomethylated and increased at the gene expression level, whereas a suppression of MuRF1/MAFbx was observed. At the protein level, we also demonstrated a significant increase in UBR5 after recovery of muscle mass from hindlimb unloading in both adult and aged rats, as well as after recovery from atrophy evoked by nerve crush injury in mice. During anabolism and hypertrophy, UBR5 gene expression increased following acute loading in three‐dimensional bioengineered mouse muscle in vitro, and after chronic electrical stimulation‐induced hypertrophy in rats in vivo, without increases in MuRF1/MAFbx. Additionally, UBR5 protein abundance increased following functional overload‐induced hypertrophy of the plantaris muscle in mice and during differentiation of primary human muscle cells. Finally, in humans, genetic association studies (>700,000 single nucleotide polymorphisms) demonstrated that the A alleles of rs10505025 and rs4734621 single nucleotide polymorphisms in the UBR5 gene were strongly associated with larger cross‐sectional area of fast‐twitch muscle fibres and favoured strength/power vs. endurance/untrained phenotypes. Overall, we suggest that: (i) UBR5 comprises a novel E3 ubiquitin ligase that is inversely regulated to MuRF1/MAFbx; (ii) UBR5 is epigenetically regulated; and (iii) UBR5 is elevated at both the gene expression and protein level during recovery from skeletal muscle atrophy and hypertrophy.

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Section: Methodsmentioning

confidence: 99%

Section: Mechanical Loading Of Bioengineered Skeletal Muscle In Vitromentioning

confidence: 99%

UBR5 is a novel E3 ubiquitin ligase involved in skeletal muscle hypertrophy and recovery from atrophy

Seaborne

Hughes

Turner

et al. 2019

The Journal of Physiology

Self Cite

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show abstract

“…Briefly, C2C12 cells were grown on T75's to approx. 80% confluence whereby 90,000 cells/ml in 2 ml growth media (GM; 4.5 g/l glucose DMEM that included 4 mM L-glutamine, 10% hiFBS, 10% hiNBCS, supplemented with an additional 2 mM L-glutamine (Girven et al, 2016),100 U/ml penicillin, 100 μ g/ml streptomycin, 0.5 mg/ml 6- 3.5% hiFBS/ 3.5% hiNBCS) maintenance media (MM) to promote myotube formation for a further 10 days.…”

Section: Mechanical Loading Of Bioengineered Skeletal Muscle In-vitromentioning

confidence: 99%

UBR5 is a Novel E3 Ubiquitin Ligase involved in Skeletal Muscle Hypertrophy and Recovery from Atrophy

Seaborne

Turner

et al. 2019

Preprint

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We aimed to investigate a novel and uncharacterised E3 ubiquitin ligase in skeletal muscle atrophy, recovery from atrophy/injury, anabolism and hypertrophy. We demonstrated an alternate gene expression profile for UBR5 versus well characterised E3-ligases, MuRF1/MAFbx, where after atrophy evoked by continuous-low-frequency electrical-stimulation in rats, MuRF1/MAFbx were both elevated yet UBR5 was unchanged. Furthermore, after recovery of muscle mass post tetrodotoxin (TTX) induced-atrophy in rats, UBR5 was hypomethylated and increased at the gene expression level, while a suppression of MuRF1/MAFbx was observed. At the protein level, we also demonstrated a significant increase in UBR5 after recovery of muscle mass from hindlimb unloading in both adult and aged rats, and after recovery from atrophy evoked by nerve crush injury in mice. During anabolism and hypertrophy, UBR5 gene expression increased following acute loading in three-dimensional bioengineered mouse muscle in-vitro, and after chronic electrical-stimulation-induced hypertrophy in rats in-vivo, without increases in MuRF1/MAFbx. Additionally, UBR5 protein abundance increased following functional overload-induced hypertrophy of the plantaris muscle in mice and during differentiation of primary human muscle cells. Finally, in humans, genetic association studies (>700,000 SNPs) demonstrated that the A alleles of rs10505025 and rs4734621 SNPs in the UBR5 gene were strongly associated with larger cross-sectional area of fast-twitch muscle fibres and favoured strength/power versus endurance/untrained phenotypes. Overall, we suggest that UBR5 is a novel E3 ubiquitin ligase that is inversely regulated to MuRF1/MAFbx, is epigenetically regulated, and is elevated at both the gene expression and protein level during recovery from skeletal muscle atrophy and hypertrophy.

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“…The physiological role of glutamine metabolism during embryonic development is not fully known [5], especially, considering the proliferation, differentiation and maturation of muscle tissue, the role of glutamine appears to be crucial. Undoubtedly, glutamine supports therapy reducing the degradation of muscle tissue, caused by inflammation, partly by satellite cell activation, especially after damaging exercise [6,7]. Interestingly, experiments with the C2C12 myoblasts' cell line, have shown that L-Glu improves skeletal muscle cell differentiation and prevents myotube atrophy after cytokine (TNF-α) via regulation of p38 MAPK [7].…”

Section: Introductionmentioning

confidence: 99%

Graphene oxide nanofilm and the addition of L-glutamine can promote development of embryonic muscle cells

Zielińska‐Górska

Hotowy

Wierzbicki

et al. 2020

Preprint

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Background: Formation of muscular pseudo-tissue depends on muscle precursor cells, the extracellular matrix (ECM)-mimicking structure and factors stimulating cell differentiation. These three things cooperate and can create a tissue-like structure, however, their interrelationships are relatively unknown. The objective was to study the interaction between surface properties, culture medium composition and heterogeneous cell culture. We would like to demonstrate that changing the surface properties by coating with graphene oxide nanofilm (nGO) can affect cell behaviour and especially their need for the key amino acid L-glutamine (L-Glu).Results: Chicken embryo muscle cells and their precursors, cultured in vitro, were used as the experimental model. The mesenchymal stem cell, collected from the hind limb of the chicken embryo at day 8 were divided into 4 groups; the control group and groups treated with nGO, L-Glu and nGO supplied with L-Glu (nGOxL-Glu). The roughness of the surface of the plastic plate covered with nGO was much lower than a standard plate. The test of nGO biocompatibility demonstrated that the cells were willing to settle on the nGO without any toxic effects. Moreover, nGO by increasing hydrophilicity and reducing roughness and presumably through chemical bonds available on the GO surface stimulated the colonisation of primary stromal cells that promote embryonic satellite cells. The viability significantly increased in cells cultured on nGOxL-Glu. Observations of cell morphology showed that the most mature state of myogenesis was characteristic for the group nGOxL-Glu. This result was confirmed by increasing the expression of MYF5 genes at mRNA and protein levels. nGO also increased the expression of MYF5 and also very strongly the expression of PAX7 at mRNA and protein levels. However, when analysing the expression of PAX7, a positive link was observed between the nGO surface and the addition of L-Glu.Conclusions: The use of nGO and L-Glu supplement may improve myogenesis and also the myogenic potential of myocytes and their precursors by promoting the formation of satellite cells. Studies have, for the first time, demonstrated positive cooperation between surface properties nGO and L-Glu supplementation to the culture medium regarding the myogenic potential of cells involved in muscle formation.

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l‐glutamine Improves Skeletal Muscle Cell Differentiation and Prevents Myotube Atrophy After Cytokine (TNF‐α) Stress Via Reduced p38 MAPK Signal Transduction

Cited by 44 publications

References 86 publications

UBR5 is a novel E3 ubiquitin ligase involved in skeletal muscle hypertrophy and recovery from atrophy

UBR5 is a novel E3 ubiquitin ligase involved in skeletal muscle hypertrophy and recovery from atrophy

UBR5 is a Novel E3 Ubiquitin Ligase involved in Skeletal Muscle Hypertrophy and Recovery from Atrophy

Graphene oxide nanofilm and the addition of L-glutamine can promote development of embryonic muscle cells

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