Degradation of ribosome and chaperone proteins is attenuated during the Differentiation of Replicatively Aged C2C12 Myoblasts

Age-related impairments in myoblast differentiation may contribute to reductions in muscle function in older adults, however, the underlying proteostasis processes are not well understood. Young (P6-10) and replicatively aged (P48-50) C2C12 myoblast cultures were investigated during early (0h-24h) and late (72h-96h) stages of differentiation using deuterium oxide (D2O) labelling and mass spectrometry. The absolute dynamic profiling technique for proteomics (Proteo-ADPT) was applied to quantify the absolute rates of abundance change, synthesis and degradation of individual proteins. Proteo-ADPT encompassed 116 proteins and 74 proteins exhibited significantly (P<0.05, FDR <5 %) different changes in abundance between young and aged cells at early and later periods of differentiation. Young cells exhibited a steady pattern of growth, protein accretion and fusion, whereas aged cells failed to gain protein mass or undergo fusion during later differentiation. Maturation of the proteome was retarded in aged myoblasts at the onset of differentiation, but their proteome appeared to "catch up" with the young cells during the early phase of the differentiation period. However, this "catch up" process in aged cells was not accomplished by higher levels of protein synthesis. Instead, a lower level of protein degradation in aged cells was responsible for the elevated gains in protein abundance. Our novel data point to a loss of proteome quality as a precursor to the lack of fusion of aged myoblasts and highlights dysregulation of protein degradation, particularly of ribosomal and chaperone proteins, as a key mechanism that may contribute to age-related declines in the capacity of myoblasts to undergo differentiation.

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Skeletal and cardiac muscle protein turnover during cold acclimation in young rats

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.2000.278.3.r705 ◽

2000 ◽

Vol 278 (3) ◽

pp. R705-R711 ◽

Cited By ~ 8

Author(s):

T. A. McAllister ◽

J. R. Thompson ◽

S. E. Samuels

Keyword(s):

Skeletal Muscle ◽

Protein Synthesis ◽

Cardiac Muscle ◽

Cold Acclimation ◽

Protein Turnover ◽

Muscle Protein ◽

Muscle Protein Turnover ◽

Protein Mass ◽

The Absolute

The effect of long-term cold exposure on skeletal and cardiac muscle protein turnover was investigated in young growing animals. Two groups of 36 male 28-day-old rats were maintained at either 5°C (cold) or 25°C (control). Rates of protein synthesis and degradation were measured in vivo on days 5, 10, 15, and 20. Protein mass by day 20 was ∼28% lower in skeletal muscle (gastrocnemius and soleus) and ∼24% higher in heart in cold compared with control rats ( P < 0.05). In skeletal muscle, the fractional rates of protein synthesis ( k syn) and degradation ( k deg) were not significantly different between cold and control rats, although k syn was lower (approximately −26%) in cold rats on day 5; consequent to the lower protein mass, the absolute rates of protein synthesis (approximately −21%; P < 0.05) and degradation (approximately −13%; P < 0.1) were lower in cold compared with control rats. In heart, overall, k syn(approximately +12%; P < 0.1) and k deg(approximately +22%; P < 0.05) were higher in cold compared with control rats; consequently, the absolute rates of synthesis (approximately +44%) and degradation (approximately +54%) were higher in cold compared with control rats ( P < 0.05). Plasma triiodothyronine concentration was higher ( P < 0.05) in cold compared with control rats. These data indicate that long-term cold acclimation in skeletal muscle is associated with the establishment of a new homeostasis in protein turnover with decreased protein mass and normal fractional rates of protein turnover. In heart, unlike skeletal muscle, rates of protein turnover did not appear to immediately return to normal as increased rates of protein turnover were observed beyond day 5. These data also indicate that increased rates of protein turnover in skeletal muscle are unlikely to contribute to increased metabolic heat production during cold acclimation.

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Identification of Withania somnifera-Silybum marianum-Trigonella foenum-graecum Formulation as a Nutritional Supplement to Contrast Muscle Atrophy and Sarcopenia

Nutrients ◽

10.3390/nu13010049 ◽

2020 ◽

Vol 13 (1) ◽

pp. 49

Author(s):

Laura Salvadori ◽

Manuela Mandrone ◽

Tommaso Manenti ◽

Catia Ercolani ◽

Luca Cornioli ◽

...

Keyword(s):

Protein Kinase ◽

Muscle Mass ◽

Muscle Atrophy ◽

Withania Somnifera ◽

Myoblast Differentiation ◽

C2c12 Myotubes ◽

Silybum Marianum ◽

Trigonella Foenum Graecum ◽

Age Related

Background: Muscle atrophy, i.e., the loss of skeletal muscle mass and function, is an unresolved problem associated with aging (sarcopenia) and several pathological conditions. The imbalance between myofibrillary protein breakdown (especially the adult isoforms of myosin heavy chain, MyHC) and synthesis, and the reduction of muscle regenerative potential are main causes of muscle atrophy. Methods: Starting from one-hundred dried hydroalcoholic extracts of medical plants, we identified those able to contrast the reduction of C2C12 myotube diameter in well-characterized in vitro models mimicking muscle atrophy associated to inflammatory states, glucocorticoid treatment or nutrient deprivation. Based on their ability to rescue type II MyHC (MyHC-II) expression in atrophying conditions, six extracts with different phytochemical profiles were selected, mixed in groups of three, and tested on atrophic myotubes. The molecular mechanism underpinning the effects of the most efficacious formulation, and its efficacy on myotubes obtained from muscle biopsies of young and sarcopenic subjects were also investigated. Results: We identified WST (Withania somnifera, Silybum marianum, Trigonella foenum-graecum) formulation as extremely efficacious in protecting C2C12 myotubes against MyHC-II degradation by stimulating Akt (protein kinase B)-dependent protein synthesis and p38 MAPK (p38 mitogen-activated protein kinase)/myogenin-dependent myoblast differentiation. WST sustains trophism in C2C12 and young myotubes, and rescues the size, developmental MyHC expression and myoblast fusion in sarcopenic myotubes. Conclusion: WST strongly counteracts muscle atrophy associated to different conditions in vitro. The future validation in vivo of our results might lead to the use of WST as a food supplement to sustain muscle mass in diffuse atrophying conditions, and to reverse the age-related functional decline of human muscles, thus improving people quality of life and reducing social and health-care costs.

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PGC-1α is associated with C2C12 Myoblast differentiation

Open Life Sciences ◽

10.2478/s11535-014-0341-y ◽

2014 ◽

Vol 9 (11) ◽

pp. 1030-1036 ◽

Cited By ~ 5

Author(s):

Yaqiu Lin ◽

Yanying Zhao ◽

Ruiwen Li ◽

Jiaqi Gong ◽

Yucai Zheng ◽

...

Keyword(s):

Mrna Level ◽

Biological Significance ◽

C2c12 Cells ◽

Myoblast Differentiation ◽

C2c12 Myoblast ◽

Mrna Levels ◽

Transfected Cells ◽

Myosin Heavy Chain Isoform ◽

Important Mediator

AbstractPGC-1α has been implicated as an important mediator of functional capacity of skeletal muscle. However, the role of PGC-1α in myoblast differentiation remains unexplored. In the present study, we observed a significant up-regulation of PGC-1α expression during the differentiation of murine C2C12 myoblast. To understand the biological significance of PGC-1α up-regulation in myoblast differentiation, C2C12 cells were transfected with murine PGC-1α cDNA and siRNA targeting PGC-1α, respectively. PGC-1α over-expressing clones fused to form typical myotubes with higher mRNA level of myosin heavy chain isoform I (MyHCI) and lower MyHCIIX. No obvious differentiation was observed in PGC-1α-targeted siRNA-transfected cells with marked decrement of mRNA levels of MyHCI and MyHCIIX. Furthermore, PGC-1α increased the expression of MyoD and MyoG in C2C12 cells, which controlled the commitment of precursor cells to myotubes. These results indicate that PGC-1α is associated with myoblast differentiation and elevates MyoD and MyoG expression levels in C2C12 cells.

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Positive feedback control between STIM1 and NFATc3 is required for C2C12 myoblast differentiation

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2012.11.082 ◽

2013 ◽

Vol 430 (2) ◽

pp. 722-728 ◽

Cited By ~ 17

Author(s):

Tam Thi Thanh Phuong ◽

Yun-Ha Yun ◽

Seon Jeong Kim ◽

Tong Mook Kang

Keyword(s):

Feedback Control ◽

Positive Feedback ◽

Myoblast Differentiation ◽

C2c12 Myoblast

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MicroRNAs Regulate Cellular ATP Levels by Targeting Mitochondrial Energy Metabolism Genes during C2C12 Myoblast Differentiation

PLoS ONE ◽

10.1371/journal.pone.0127850 ◽

2015 ◽

Vol 10 (5) ◽

pp. e0127850 ◽

Cited By ~ 23

Author(s):

Puntita Siengdee ◽

Nares Trakooljul ◽

Eduard Murani ◽

Manfred Schwerin ◽

Klaus Wimmers ◽

...

Keyword(s):

Energy Metabolism ◽

Myoblast Differentiation ◽

C2c12 Myoblast ◽

Mitochondrial Energy Metabolism ◽

Atp Levels

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Catch-Up Growth in Children Born Small for Gestational Age Related to Body Composition and Metabolic Risk at Six Years of Age in the UK

Hormone Research in Paediatrics ◽

10.1159/000508974 ◽

2020 ◽

Vol 93 (2) ◽

pp. 119-127

Author(s):

M. Loredana Marcovecchio ◽

Samantha Gorman ◽

Laura P.E. Watson ◽

David B. Dunger ◽

Kathryn Beardsall

Keyword(s):

Body Composition ◽

Gestational Age ◽

Small For Gestational Age ◽

Metabolic Risk ◽

Age Related ◽

Catch Up ◽

The Uk

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Proteomics-based investigation in C2C12 myoblast differentiation

European Journal of Histochemistry ◽

10.4081/ejh.2009.e31 ◽

2009 ◽

Vol 53 (4) ◽

pp. 31 ◽

Cited By ~ 11

Author(s):

L. Casadei ◽

L. Vallorani ◽

A.M. Gioacchini ◽

M. Guescini ◽

S. Burattini ◽

...

Keyword(s):

Myoblast Differentiation ◽

C2c12 Myoblast

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Myoferlin Regulates Wnt/β-Catenin Signaling-Mediated Skeletal Muscle Development by Stabilizing Dishevelled-2 Against Autophagy

International Journal of Molecular Sciences ◽

10.3390/ijms20205130 ◽

2019 ◽

Vol 20 (20) ◽

pp. 5130 ◽

Cited By ~ 1

Author(s):

Shunshun Han ◽

Can Cui ◽

Haorong He ◽

Xiaoxu Shen ◽

Yuqi Chen ◽

...

Keyword(s):

Skeletal Muscle ◽

Muscle Atrophy ◽

Muscle Development ◽

Skeletal Muscle Atrophy ◽

Myoblast Differentiation ◽

C2c12 Myoblast ◽

Skeletal Muscle Development ◽

Muscle Tissues ◽

Underlying Mechanisms ◽

Canonical Wnt

Myoferlin (MyoF), which is a calcium/phospholipid-binding protein expressed in cardiac and muscle tissues, belongs to the ferlin family. While MyoF promotes myoblast differentiation, the underlying mechanisms remain poorly understood. Here, we found that MyoF not only promotes C2C12 myoblast differentiation, but also inhibits muscle atrophy and autophagy. In the present study, we found that myoblasts fail to develop into mature myotubes due to defective differentiation in the absence of MyoF. Meanwhile, MyoF regulates the expression of atrophy-related genes (Atrogin-1 and MuRF1) to rescue muscle atrophy. Furthermore, MyoF interacts with Dishevelled-2 (Dvl-2) to activate canonical Wnt signaling. MyoF facilitates Dvl-2 ubiquitination resistance by reducing LC3-labeled Dvl-2 levels and antagonizing the autophagy system. In conclusion, we found that MyoF plays an important role in myoblast differentiation during skeletal muscle atrophy. At the molecular level, MyoF protects Dvl-2 against autophagy-mediated degradation, thus promoting activation of the Wnt/β-catenin signaling pathway. Together, our findings suggest that MyoF, through stabilizing Dvl-2 and preventing autophagy, regulates Wnt/β-catenin signaling-mediated skeletal muscle development.

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