Interleukin-6 promotes myogenic differentiation of mouse skeletal muscle cells: role of the STAT3 pathway

2013 ◽  
Vol 304 (2) ◽  
pp. C128-C136 ◽  
Author(s):  
Miriam Hoene ◽  
Heike Runge ◽  
Hans Ulrich Häring ◽  
Erwin D. Schleicher ◽  
Cora Weigert
Keyword(s):  
Skeletal Muscle ◽  
Mrna Expression ◽  
Myogenic Differentiation ◽  
Muscle Cells ◽  
C2c12 Cells ◽  
Skeletal Muscle Cells ◽  
Wild Type ◽  
C2c12 Myoblasts ◽  
Sequence Of Events ◽  
Primary Mouse

Myogenic differentiation of skeletal muscle cells is characterized by a sequence of events that include activation of signal transducer and activator of transcription 3 (STAT3) and enhanced expression of its target gene Socs3. Autocrine effects of IL-6 may contribute to the activation of the STAT3-Socs3 cascade and thus to myogenic differentiation. The importance of IL-6 and STAT3 for the differentiation process was studied in C2C12 cells and in primary mouse wild-type and IL-6−/− skeletal muscle cells. In differentiating C2C12 myoblasts, the upregulation of IL-6 mRNA expression and protein secretion started after increased phosphorylation of STAT3 on tyrosine 705 and increased mRNA expression of Socs3 was observed. Knockdown of STAT3 and IL-6 mRNA in differentiating C2C12 myoblasts impaired the expression of the myogenic markers myogenin and MyHC IIb and subsequently myotube fusion. However, the knockdown of IL-6 did not prevent the induction of STAT3 tyrosine phosphorylation. The IL-6-independent activation of STAT3 was verified in differentiating primary IL-6−/− myoblasts. The phosphorylation of STAT3 and the expression levels of STAT3, Socs3, and myogenin during differentiation were comparable in the primary myoblasts independent of the genotype. However, IL-6−/− cells failed to induce MyHC IIb expression to the same level as in wild-type cells and showed reduced myotube formation. Supplementation of IL-6 could partially restore the fusion of IL-6−/− cells. These data demonstrate that IL-6 depletion during myogenic differentiation does not reduce the activation of the STAT3-Socs3 cascade, while IL-6 and STAT3 are both necessary to promote myotube fusion.

scholarly journals Regulation of Lin28a-miRNA let-7b-5p pathway in skeletal muscle cells by peroxisome proliferator-activated receptor delta

2020 ◽  
Vol 319 (3) ◽  
pp. C541-C551
Author(s):  
Hygor N. Araujo ◽  
Tanes I. Lima ◽  
Dimitrius Santiago P. S. F. Guimarães ◽  
Andre G. Oliveira ◽  
Bianca C. Favero-Santos ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Energy Homeostasis ◽  
Muscle Cells ◽  
C2c12 Cells ◽  
Skeletal Muscle Cells ◽  
Peroxisome Proliferator ◽  
Peroxisome Proliferator Activated Receptor ◽  
Primary Mouse ◽  
Murine Skeletal Muscle ◽  
Muscle Biopsies

Lin28a/miRNA let-7b-5p pathway has emerged as a key regulators of energy homeostasis in the skeletal muscle. However, the mechanism through which this pathway is regulated in the skeletal muscle has remained unclear. We have found that 8 wk of aerobic training (Tr) markedly decreased let-7b-5p expression in murine skeletal muscle, whereas high-fat diet (Hfd) increased its expression. Conversely, Lin28a expression, a well-known inhibitor of let-7b-5p, was induced by Tr and decreased by Hfd. Similarly, in human muscle biopsies, Tr increased LIN28 expression and decreased let-7b-5p expression. Bioinformatics analysis of LIN28a DNA sequence revealed that its enrichment in peroxisome proliferator-activated receptor delta (PPARδ) binding sites, which is a well-known metabolic regulator of exercise. Treatment of primary mouse skeletal muscle cells or C2C12 cells with PPARδ activators GW501516 and AICAR increased Lin28a expression. Lin28a and let-7b-5p expression was also regulated by PPARδ coregulators. While PPARγ coactivator-1α (PGC1α) increased Lin28a expression, corepressor NCoR1 decreased its expression. Furthermore, PGC1α markedly reduced the let-7b-5p expression. PGC1α-mediated induction of Lin28a expression was blocked by the PPARδ inhibitor GSK0660. In agreement, Lin28a expression was downregulated in PPARδ knocked-down cells leading to increased let-7b-5p expression. Finally, we show that modulation of the Lin28a- let-7b-5p pathway in muscle cells leads to changes in mitochondrial metabolism in PGC1α dependent fashion. In summary, we demonstrate that Lin28a- let-7b-5p is a direct target of PPARδ in the skeletal muscle, where it impacts mitochondrial respiration.

scholarly journals Changes of Myogenic Reactive Oxygen Species and Interleukin-6 in Contracting Skeletal Muscle Cells

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Hongying Pan ◽  
Xiaoyang Xu ◽  
Xuanming Hao ◽  
Yajun Chen
Keyword(s):  
Reactive Oxygen Species ◽  
Skeletal Muscle ◽  
Mrna Expression ◽  
Reactive Oxygen ◽  
Muscle Cells ◽  
C2c12 Cells ◽  
Skeletal Muscle Cells ◽  
C2c12 Myotubes ◽  
Ros Generation ◽  
Oxygen Species

The aim of this study was to measure changes in myotube reactive oxygen species (ROS) and the production of interleukin (IL)-6 in electrically stimulated mouse C2C12 skeletal muscle cells. After five days of differentiation, myotubes were stimulated using an electrical stimulator set at 45 V at a frequency of 5 Hz, with a pulse width of 20 ms. Acute stimulations were performed for 45, 60, 75, 90, or 120 min in each dish. ROSs were detected in the extracted cells directly using a fluorescent probe. IL-6 mRNA expression in C2C12 myotubes and IL-6 concentration in C2C12 myotube supernatants were determined using real-time PCR and ELISA, respectively. Compared with control cells, ROS generation was significantly increased at 45 min after the onset of stimulation (P<0.01) and continued to increase, reaching a maximum at 120 min. IL-6 mRNA expression and IL-6 concentration in C2C12 cells were significantly increased after 75 min (P<0.01) and 120 min (P<0.05) of electrical stimulation (ES) compared with the control cells. Our data show that a specific ES intensity may modulate ROS accumulation and affect IL-6 gene expression in contracting skeletal muscle cells.

scholarly journals The potassium channels TASK2 and TREK1 regulate functional differentiation of murine skeletal muscle cells

2016 ◽  
Vol 311 (4) ◽  
pp. C583-C595 ◽  
Author(s):  
Ali M. Afzali ◽  
Tobias Ruck ◽  
Alexander M. Herrmann ◽  
Janette Iking ◽  
Claudia Sommer ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Muscle Tissue ◽  
Outward Current ◽  
Muscle Cells ◽  
Human Muscle ◽  
C2c12 Cells ◽  
Skeletal Muscle Cells ◽  
Mouse Muscle ◽  
Organ Systems ◽  
Primary Mouse

Two-pore domain potassium (K2P) channels influence basic cellular parameters such as resting membrane potential, cellular excitability, or intracellular Ca2+-concentration [Ca2+]i. While the physiological importance of K2P channels in different organ systems (e.g., heart, central nervous system, or immune system) has become increasingly clear over the last decade, their expression profile and functional role in skeletal muscle cells (SkMC) remain largely unknown. The mouse SkMC cell line C2C12, wild-type mouse muscle tissue, and primary mouse muscle cells (PMMs) were analyzed using quantitative PCR, Western blotting, and immunohistochemical stainings as well as functional analysis including patch-clamp measurements and Ca2+ imaging. Mouse SkMC express TWIK-related acid-sensitive K+ channel (TASK) 2, TWIK-related K+ channel (TREK) 1, TREK2, and TWIK-related arachidonic acid stimulated K+ channel (TRAAK). Except TASK2 all mentioned channels were upregulated in vitro during differentiation from myoblasts to myotubes. TASK2 and TREK1 were also functionally expressed and upregulated in PMMs isolated from mouse muscle tissue. Inhibition of TASK2 and TREK1 during differentiation revealed a morphological impairment of myoblast fusion accompanied by a downregulation of maturation markers. TASK2 and TREK1 blockade led to a decreased K+ outward current and a decrease of ACh-dependent Ca2+ influx in C2C12 cells as potential underlying mechanisms. K2P-channel expression was also detected in human muscle tissue by immunohistochemistry pointing towards possible relevance for human muscle cell maturation and function. In conclusion, our findings for the first time demonstrate the functional expression of TASK2 and TREK1 in muscle cells with implications for differentiation processes warranting further investigations in physiologic and pathophysiologic scenarios.

α-amyrin induce GLUT4 translocation mediated by AMPK and PPARδ/γ in C2C12 myoblasts

2021 ◽  
Author(s):  
Abraham Giacoman-Martínez ◽  
Francisco Javier Alarcón-Aguilar ◽  
Alejandro Zamilpa-Alvarez ◽  
Fengyang Huang ◽  
Rodrigo Romero ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Protein Kinase ◽  
Glucose Transporter ◽  
Metabolic Diseases ◽  
Muscle Cells ◽  
Skeletal Muscle Cells ◽  
Peroxisome Proliferator ◽  
Glut4 Translocation ◽  
Pentacyclic Triterpene ◽  
C2c12 Myoblasts

α-amyrin, a natural pentacyclic triterpene, have anti-hyperglycemic effect in mice and dual PPARδ/γ action in 3T3-L1 adipocytes, and potential in the control of type 2 diabetes (T2D). About 80% of glucose uptake occurs in skeletal muscle cells, playing a significant role in IR and T2D. Peroxisome-proliferator activated receptors (PPARs), in particular PPARδ and PPARγ, are involved in the regulation of lipids and carbohydrates and, along adenosine-monophosphate (AMP)-activated protein kinase (AMPK) and protein kinase B (Akt/PKB), are implicated in translocation of glucose transporter 4 (GLUT4). However, it is still unknown whether α-amyrin can affect these pathways in skeletal muscle cells. The work's objective was to determine the action of α-amyrin in PPARδ, PPARγ, AMPK, and Akt/PKB in C2C12 myoblasts. The expression of PPARδ, PPARγ, FATP, and GLUT4 was quantified using RT-qPCR and Western blot. α-amyrin increased these markers along with p-AMPK but not p-Akt/PKB. Molecular docking showed that α-amyrin acts as an AMPK-allosteric activator, and may be related to GLUT4 translocation, evidenced by confocal microscopy. These data support that α-amyrin could have an insulin-mimetic action in C2C12 myoblasts and should be considered as a bioactive molecule for new multitarget drugs with utility in T2D and other metabolic diseases.

scholarly journals 17β-Estradiol abrogates apoptosis in murine skeletal muscle cells through estrogen receptors: role of the phosphatidylinositol 3-kinase/Akt pathway

2007 ◽  
Vol 196 (2) ◽  
pp. 385-397 ◽  
Author(s):  
Andrea Vasconsuelo ◽  
Lorena Milanesi ◽  
Ricardo Boland
Keyword(s):  
Skeletal Muscle ◽  
Protective Action ◽  
Muscle Cells ◽  
Cellular Systems ◽  
C2c12 Cells ◽  
Skeletal Muscle Cells ◽  
Akt Pathway ◽  
Akt Activation ◽  
17Β Estradiol ◽  
Murine Skeletal Muscle

Estrogens can regulate apoptosis in various cellular systems. The present study shows that 17β-estradiol (E2), at physiological concentrations, abrogates DNA damage, poly (ADP-ribose) polymerase cleavage, and mitochondrial cytochrome c release induced by H2O2 or etoposide in mouse skeletal muscle C2C12 cells. This protective action, which involved PI3K/Akt activation and Bcl-2 associated death agonist (BAD) phosphorylation, was inhibited by antibodies against the estrogen receptor (ER) α or β isoforms, or transfecting siRNA specific for each isoform. The inhibition of the antiapoptotic action of E2 at the mitochondrial level was more pronounced when ER-β was immunoneutralized or suppressed by mRNA silencing, whereas transfection of C2C12 cells with either ER-α siRNA or ER-β siRNA blocked the activation of Akt by E2, suggesting differential involvement of ER isoforms depending on the step of the apoptotic/survival pathway evaluated. These results indicate that E2 exerts antiapoptotic effects in skeletal muscle cells which are mediated by ER-β and ER-α and involve the PI3K/Akt pathway.

NFAT activation by membrane potential follows a calcium pathway distinct from other activity-related transcription factors in skeletal muscle cells

2008 ◽  
Vol 294 (3) ◽  
pp. C715-C725 ◽  
Author(s):  
Juan Antonio Valdés ◽  
Eduardo Gaggero ◽  
Jorge Hidalgo ◽  
Nancy Leal ◽  
Enrique Jaimovich ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Transcription Factors ◽  
Electrical Stimulation ◽  
Stimulus Frequency ◽  
Muscle Cells ◽  
C2c12 Cells ◽  
Skeletal Muscle Cells ◽  
Mrna Levels ◽  
Activated T Cells ◽  
Nfat Activation

Depolarization of skeletal muscle cells triggers intracellular Ca2+ signals mediated by ryanodine and inositol 1,4,5-trisphosphate (IP3) receptors. Previously, we have reported that K+-induced depolarization activates transcriptional regulators ERK, cAMP response element-binding protein, c- fos, c- jun, and egr-1 through IP3-dependent Ca2+ release, whereas NF-κB activation is elicited by both ryanodine and IP3 receptor-mediated Ca2+ signals. We have further shown that field stimulation with electrical pulses results in an NF-κB activation increase dependent of the amount of pulses and independent of their frequency. In this work, we report the results obtained for nuclear factor of activated T cells (NFAT)-mediated transcription and translocation generated by both K+ and electrical stimulation protocols in primary skeletal muscle cells and C2C12 cells. The Ca2+ source for NFAT activation is through release by ryanodine receptors and extracellular Ca2+ entry. We found this activation to be independent of the number of pulses within a physiological range of stimulus frequency and enhanced by long-lasting low-frequency stimulation. Therefore, activation of the NFAT signaling pathway differs from that of NF-κB and other transcription factors. Calcineurin enzyme activity correlated well with the relative activation of NFAT translocation and transcription using different stimulation protocols. Furthermore, both K+-induced depolarization and electrical stimulation increased mRNA levels of the type 1 IP3 receptor mediated by calcineurin activity, which suggests that depolarization may regulate IP3 receptor transcription. These results confirm the presence of at least two independent pathways for excitation-transcription coupling in skeletal muscle cells, both dependent on Ca2+ release and triggered by the same voltage sensor but activating different intracellular release channels.

Altered PPARγ expression inhibits myogenic differentiation in C2C12 skeletal muscle cells

2006 ◽  
Vol 294 (1-2) ◽  
pp. 163-171 ◽  
Author(s):  
Jaskirat Singh ◽  
Navin Kumar Verma ◽  
Sejal M. Kansagra ◽  
Bhusan N. Kate ◽  
Chinmoy Sankar Dey
Keyword(s):  
Skeletal Muscle ◽  
Myogenic Differentiation ◽  
Muscle Cells ◽  
Skeletal Muscle Cells ◽  
C2c12 Skeletal Muscle Cells ◽  
Pparγ Expression

scholarly journals Alignment of Skeletal Muscle Cells Cultured in Collagen Gel by Mechanical and Electrical Stimulation

2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Takara Tanaka ◽  
Noriko Hattori-Aramaki ◽  
Ayano Sunohara ◽  
Keisuke Okabe ◽  
Yoshiaki Sakamoto ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Tissue Engineering ◽  
Electrical Stimulation ◽  
Collagen Gel ◽  
Muscle Cells ◽  
Mechanical Force ◽  
C2c12 Cells ◽  
Skeletal Muscle Cells ◽  
Collagen Gels ◽  
Cells Cultured

For in vitro tissue engineering of skeletal muscle, alignment and fusion of the cultured skeletal muscle cells are required. Although the successful alignment of skeletal muscle cells cultured in collagen gel has been reported using a mechanical force, other means of aligning cultured skeletal muscle cells have not been described. However, skeletal muscle cells cultured in a two-dimensional dish have been reported to align in a uniform direction when electrically stimulated. The purpose of this study is to determine if skeletal muscle cells cultured three-dimensionally in collagen gels can be aligned by an electrical load. By adding direct current to cells of the C2C12 skeletal muscle cell line cultured in collagen gel, it was possible to align C2C12 cells in a similar direction. However, the ratio of alignment was better when mechanical force was used as the means of alignment. Thus for tissue engineering of skeletal muscle cells, electrical stimulation may be useful as a supplementary method.

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