scholarly journals Biomechanical signals upregulate myogenic gene induction in the presence or absence of inflammation

2007 ◽  
Vol 293 (1) ◽  
pp. C267-C276 ◽  
Author(s):  
Ravi Chandran ◽  
Thomas J. Knobloch ◽  
Mirela Anghelina ◽  
Sudha Agarwal
Keyword(s):  
Cell Differentiation ◽  
Muscle Cell ◽  
Kinase Inhibitor ◽  
Cell Damage ◽  
Myogenic Differentiation ◽  
C2c12 Cell ◽  
Gene Induction ◽  
C2c12 Cells ◽  
Proinflammatory Mediators ◽  
Tnf Α

Inflammation of the muscle invariably leads to muscle cell damage and impaired regeneration. Biomechanical signals play a vital role in the regulation of myogenesis in healthy and inflamed muscle. We hypothesized that biomechanical signals counteract the actions of proinflammatory mediators and upregulate the basic helix-loop-helix and MADS box transcription enhancer factor 2 (MEF2) families of transcription factors, leading to increased myogenesis in inflamed muscle cells. For this purpose, C2C12 cells plated on collagenized silastic membranes were subjected to equibiaxial cyclic tensile strain (CTS) in the presence or absence of TNF-α, and the myogenic gene induction was examined over a period of 72 h. Exposure of cells to CTS resulted in a significant upregulation of mRNA expressions and synthesis of myogenic regulatory factors, MYOD1, myogenin (MYOG), MEF2A, and cyclin-dependent kinase inhibitor 1A (CDKN1A; p21) as well as muscle structural proteins like myosin heavy chain (MYHC) isoforms (MYH1, MYH2, and MYH4) and α-tropomyosin (TPM1), eventually leading to an increase in myotube formation. Contrarily, TNF-α suppressed the expression of all of the above differentiation-inducing factors in C2C12 cells. Further results revealed that simultaneous exposure of C2C12 cells to CTS and TNF-α abrogated the TNF-α-mediated downregulation of myogenic differentiation. In fact, the mRNA expression and protein synthesis of all myogenic factors ( Myod1, Myog, Mef2a, Cdkn1a, Myh1, Myh2, Myh4, and Tpm1) were increased in stretched C2C12 cells despite the sustained presence of TNF-α. These results demonstrate that mechanotransduction regulates multiple signaling molecules involved in C2C12 cell differentiation. On one hand, these signals are potent transducers of myotube phenotype in myoblasts; on the other, these signals counteract catabolic actions of proinflammatory cytokines like TNF-α and allow the expression of myogenic genes to upregulate muscle cell differentiation.

scholarly journals Doxycycline Inhibits IL-17-Stimulated MMP-9 Expression by Downregulating ERK1/2 Activation: Implications in Myogenic Differentiation

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Hristina Obradović ◽  
Jelena Krstić ◽  
Tamara Kukolj ◽  
Drenka Trivanović ◽  
Ivana Okić Đorđević ◽  
...  
Keyword(s):  
Inflammatory Diseases ◽  
Myogenic Differentiation ◽  
C2c12 Cell ◽  
C2c12 Cells ◽  
Myoblast Differentiation ◽  
Interleukin 17 ◽  
Myoblast Cells ◽  
Pathological Consequence ◽  
Muscle Remodeling

Interleukin 17 (IL-17) is a cytokine with pleiotropic effects associated with several inflammatory diseases. Although elevated levels of IL-17 have been described in inflammatory myopathies, its role in muscle remodeling and regeneration is still unknown. Excessive extracellular matrix degradation in skeletal muscle is an important pathological consequence of many diseases involving muscle wasting. In this study, the role of IL-17 on the expression of matrix metalloproteinase- (MMP-) 9 in myoblast cells was investigated. The expression of MMP-9 after IL-17 treatment was analyzed in mouse myoblasts C2C12 cell line. The increase in MMP-9 production by IL-17 was concomitant with its capacity to inhibit myogenic differentiation of C2C12 cells. Doxycycline (Doxy) treatment protected the myogenic capacity of myoblasts from IL-17 inhibition and, moreover, increased myotubes hypertrophy. Doxy blocked the capacity of IL-17 to stimulate MMP-9 production by regulating IL-17-induced ERK1/2 MAPK activation. Our results imply that MMP-9 mediates IL-17’s capacity to inhibit myoblast differentiation during inflammatory diseases and indicate that Doxy can modulate myoblast response to inflammatory induction by IL-17.

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

2016 ◽  
Vol 231 (12) ◽  
pp. 2720-2732 ◽  
Author(s):  
Matthew Girven ◽  
Hannah F. Dugdale ◽  
Daniel J. Owens ◽  
David C. Hughes ◽  
Claire E. Stewart ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Signal Transduction ◽  
Cell Differentiation ◽  
P38 Mapk ◽  
Muscle Cell ◽  
Skeletal Muscle Cell ◽  
Muscle Cell Differentiation ◽  
Tnf Α

scholarly journals TNF-α and IGF1 modify the microRNA signature in skeletal muscle cell differentiation

2015 ◽  
Vol 13 (1) ◽  
pp. 4 ◽  
Author(s):  
Swanhild U Meyer ◽  
Christian Thirion ◽  
Anna Polesskaya ◽  
Stefan Bauersachs ◽  
Sebastian Kaiser ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Cell Differentiation ◽  
Muscle Cell ◽  
Skeletal Muscle Cell ◽  
Muscle Cell Differentiation ◽  
Tnf Α

scholarly journals Coupled Transcriptional and Translational Control of Cyclin-Dependent Kinase Inhibitor p18INK4cExpression during Myogenesis

1998 ◽  
Vol 18 (4) ◽  
pp. 2334-2343 ◽  
Author(s):  
Dawn E. Phelps ◽  
Kuang-Ming Hsiao ◽  
Yan Li ◽  
Nanpin Hu ◽  
David S. Franklin ◽  
...  
Keyword(s):  
Translational Control ◽  
Kinase Inhibitor ◽  
Myogenic Differentiation ◽  
Protein A ◽  
Terminal Differentiation ◽  
C2c12 Cells ◽  
Cdk Inhibitor ◽  
Cyclin Dependent Kinase ◽  
C2c12 Myoblasts ◽  
Exon 1

ABSTRACT Terminal differentiation of many cell types involves permanent withdrawal from the cell division cycle. The p18 INK4c protein, a member of the p16/INK4 cyclin-dependent kinase (CDK) inhibitor family, is induced more than 50-fold during myogenic differentiation of mouse C2C12 myoblasts to become the predominant CDK inhibitor complexed with CDK4 and CDK6 in terminally differentiated myotubes. We have found that the p18 INK4c gene expresses two mRNA transcripts—a 2.4-kb transcript, p18(L), and a 1.2-kb transcript, p18(S). In proliferating C2C12 myoblasts, only the larger p18(L) transcript is expressed from an upstream promoter. As C2C12 cells are induced to differentiate into permanently arrested myotubes, the abundance of the p18(L) transcript decreases. The smaller p18(S) transcript expressed from a downstream promoter becomes detectable by 12 h postinduction and is the predominant transcript expressed in terminally differentiated myotubes. Both transcripts contain coding exons 2 and 3, but p18(L) uniquely contains an additional noncoding 1.2-kb exon, exon 1, corresponding exclusively to the 5′ untranslated region (5′ UTR). The expression pattern of the shorter p18(S) transcript, but not that of the longer p18(L) transcript, correlates with terminal differentiation of muscle, lung, liver, thymus, and eye lens cells during mouse embryo development. The presence of the long 5′ UTR in exon 1 attenuated the translation of p18(L) transcript, while its absence from the shorter p18(S) transcript resulted in significantly more efficient translation of the p18 protein. Our results demonstrate that during terminal muscle cell differentiation, induction of the p18 protein is regulated by promoter switching coupled with translational control.

scholarly journals A Role of Alkbh5 at the Early Stage of Cell Differentiation in C2C12 Cells

2021 ◽  
Vol 5 (Supplement_2) ◽  
pp. 1204-1204
Author(s):  
Ju Yeong Cho ◽  
Seo Yoon Choi ◽  
Moon Jung Choi ◽  
Yoon Jung Park
Keyword(s):  
Cell Differentiation ◽  
Early Stage ◽  
Myogenic Differentiation ◽  
Rna Isolation ◽  
C2c12 Cells ◽  
Cancer Center ◽  
Translation Efficiency ◽  
Dot Blot ◽  
Funding Sources ◽  
Epigenetic Modifier

Abstract Objectives N6-methyladenosine(m6A) is one of the most abundant mRNA modifications, controlling mRNA metabolism including splicing, stabilization, and translation efficiency. Although it has been reported that m6A was involved in cellular differentiation process, its underlying molecular mechanism is not fully understood. The object of this study was to discover ALKBH5, a m6A demethylase, as a key regulator in early myogenesis and adipogenesis using C2C12 model. Methods To identify the epigenetic modifier, we compared publicly available GEO data sets from two independent adipogenic and myogenic differentiation models. M6A levels of mRNAs were measured by dot blot assays upon differentiation. C2C12 cells were maintained under myogenesis or adipogenesis media(±rosiglitazone) for 5–8 days. Alkbh5 knockdown using siRNA and overexpression using GFP-tagged cDNA plasmid were transfected into cells 2 days before confluence. Gene expression analysis was done by qRT-PCR, following RNA isolation and conversion into cDNA. Oil-red-O staining was conducted to detect lipid droplets. Myotube length and width were measured by giemsa staining. Results We found Alkbh5 as a common epigenetic modifier, expression of which was significantly different in both early myogenic and adipogenic cells, compared to undifferentiated cells. Consistently, m6A levels, measured by dot blots, decreased during myogenesis(day 2), while increased during adipogenesis(24h). To examine if modulation of Alkbh5 expression affected cell differentiation, we conducted both Alkbh5 knockdown and overexpression in C2C12 myoblasts, followed by differentiating stimulation into myotube and adipocyte by appropriate media. Knockdown of Alkbh5 facilitates lipid accumulation, while overexpression of Alkbh5 represents significant increase in myotube length and width. Expression of myogenesis and adipogenesis markers were consistent with the phenotypes. Conclusions These results suggested that Alkbh5 had an important role in determination of cell fates towards myogenesis over adipogenesis at the early point of cell differentiation. Funding Sources This study was supported by the National Research Foundation of Korea the Korean National Cancer Center. JYC was supported by Brain Korea Four Project(Education Research Center for 4IR-Based Health Care).

scholarly journals p27Kip1 Acts Downstream of N-Cadherin-mediated Cell Adhesion to Promote Myogenesis beyond Cell Cycle Regulation

2005 ◽  
Vol 16 (3) ◽  
pp. 1469-1480 ◽  
Author(s):  
Graziella Messina ◽  
Cristiana Blasi ◽  
Severina Anna La Rocca ◽  
Monica Pompili ◽  
Attilio Calconi ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Myogenic Differentiation ◽  
Cell Cycle Control ◽  
C2c12 Cell ◽  
Terminal Differentiation ◽  
Active Role ◽  
C2c12 Cells ◽  
Low Density

It is widely acknowledged that cultured myoblasts can not differentiate at very low density. Here we analyzed the mechanism through which cell density influences myogenic differentiation in vitro. By comparing the behavior of C2C12 myoblasts at opposite cell densities, we found that, when cells are sparse, failure to undergo terminal differentiation is independent from cell cycle control and reflects the lack of p27Kip1 and MyoD in proliferating myoblasts. We show that inhibition of p27Kip1 expression impairs C2C12 cell differentiation at high density, while exogenous p27Kip1 allows low-density cultured C2C12 cells to enter the differentiative program by regulating MyoD levels in undifferentiated myoblasts. We also demonstrate that the early induction of p27Kip1 is a critical step of the N-cadherin-dependent signaling involved in myogenesis. Overall, our data support an active role of p27Kip1 in the decision of myoblasts to commit to terminal differentiation, distinct from the regulation of cell proliferation, and identify a pathway that, reasonably, operates in vivo during myogenesis and might be part of the phenomenon known as “community effect”.

scholarly journals Fibroblast Growth Factor 21 Promotes C2C12 Cells Myogenic Differentiation by Enhancing Cell Cycle Exit

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Xinyi Liu ◽  
Yongliang Wang ◽  
Shuhong Zhao ◽  
Xinyun Li
Keyword(s):  
Cell Cycle ◽  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Myogenic Differentiation ◽  
C2c12 Cell ◽  
C2c12 Cells ◽  
Fibroblast Growth Factor 21 ◽  
Fibroblast Growth ◽  
Cell Cycle Exit ◽  
Protein Functions

Fibroblast growth factor 21 (FGF21), a secretion protein, functions as a pivotal regulator of energy metabolism and is being considered as a therapeutic candidate in metabolic syndromes. However, the roles of FGF21 in myogenic differentiation and cell cycle remain obscure. In this study, we investigated the function of FGF21 in myogenesis and cell cycle exit using C2C12 cell line. Our data showed that the expression of myogenic genes as well as cell cycle exit genes was increased after FGF21 overexpression, and FGF21 overexpression induces cell cycle arrest. Moreover, cell cycle genes were decreased in FGF21 overexpression cells while they were increased in FGF21 knockdown cells. Further, FGF21/P53/p21/Cyclin-CDK has been suggested as the key pathway for cell cycle exit mediated by FGF21 in C2C12 cells. Also, we deduce that FGF21 promotes the initiation of myogenic differentiation mainly through enhancing cell cycle exit of C2C12 cells. Taken together, our results demonstrated that FGF21 promotes cell cycle exit and enhances myogenic differentiation of C2C12 cells. This study provided new evidence that FGF21 promotes myogenic differentiation, which could be useful for better understanding the roles of FGF21 in myogenesis.

scholarly journals Role of Transmembrane 4 Superfamily (Tm4sf) Proteins Cd9 and Cd81 in Muscle Cell Fusion and Myotube Maintenance

1999 ◽  
Vol 146 (4) ◽  
pp. 893-904 ◽  
Author(s):  
Isao Tachibana ◽  
Martin E. Hemler
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Fusion ◽  
Muscle Cell ◽  
C2c12 Cell ◽  
Ectopic Expression ◽  
C2c12 Cells ◽  
C2c12 Myoblast ◽  
Syncytia Formation ◽  
Transmembrane 4 Superfamily

The role of transmembrane 4 superfamily (TM4SF) proteins during muscle cell fusion has not been investigated previously. Here we show that the appearance of TM4SF protein, CD9, and the formation of CD9–β1 integrin complexes were both regulated in coordination with murine C2C12 myoblast cell differentiation. Also, anti-CD9 and anti-CD81 monoclonal antibodies substantially inhibited and delayed conversion of C2C12 cells to elongated myotubes, without affecting muscle-specific protein expression. Studies of the human myoblast-derived RD sarcoma cell line further demonstrated that TM4SF proteins have a role during muscle cell fusion. Ectopic expression of CD9 caused a four- to eightfold increase in RD cell syncytia formation, whereas anti-CD9 and anti-CD81 antibodies markedly delayed RD syncytia formation. Finally, anti-CD9 and anti-CD81 monoclonal antibodies triggered apoptotic degeneration of C2C12 cell myotubes after they were formed. In summary, TM4SF proteins such as CD9 and CD81 appear to promote muscle cell fusion and support myotube maintenance.

scholarly journals Hes6 regulates myogenic differentiation

Development ◽  
2002 ◽  
Vol 129 (9) ◽  
pp. 2195-2207
Author(s):  
Judy Cossins ◽  
Ann E. Vernon ◽  
Yun Zhang ◽  
Anna Philpott ◽  
Philip H. Jones
Keyword(s):  
Protein Interactions ◽  
Kinase Inhibitor ◽  
Myogenic Differentiation ◽  
C2c12 Cells ◽  
Myoblast Differentiation ◽  
C2c12 Myoblast ◽  
Xenopus Embryos ◽  
Enhancer Of Split

Hes6 is a basic helix-loop-helix transcription factor homologous to Drosophila Enhancer of Split (EoS) proteins. It is known to promote neural differentiation and to bind to Hes1, a related protein that is part of the Notch signalling pathway, affecting Hes1-regulated transcription. We show that Hes6 is expressed in the murine embryonic myotome and is induced on C2C12 myoblast differentiation in vitro. Hes6 binds DNA containing the Enhancer of Split E box (ESE) motif, the preferred binding site of Drosophila EoS proteins, and represses transcription of an ESE box reporter. When overexpressed in C2C12 cells, Hes6 impairs normal differentiation, causing a decrease in the induction of the cyclin-dependent kinase inhibitor, p21Cip1, and an increase in the number of cells that can be recruited back into the cell cycle after differentiation in culture. In Xenopus embryos, Hes6 is co-expressed with MyoD in early myogenic development. Microinjection of Hes6 RNA in vivo in Xenopus embryos results in an expansion of the myotome, but suppression of terminal muscle differentiation and disruption of somite formation at the tailbud stage. Analysis of Hes6 mutants indicates that the DNA-binding activity of Hes6 is not essential for its myogenic phenotype, but that protein-protein interactions are. Thus, we demonstrate a novel role for Hes6 in multiple stages of muscle formation.

scholarly journals Mdfi Promotes C2C12 Cell Differentiation and Positively Modulates Fast-to-Slow-Twitch Muscle Fiber Transformation

2021 ◽  
Vol 9 ◽  
Author(s):  
Bo Huang ◽  
Yiren Jiao ◽  
Yifan Zhu ◽  
Zuocheng Ning ◽  
Zijian Ye ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Muscle Fiber ◽  
Fiber Type ◽  
C2c12 Cell ◽  
C2c12 Cells ◽  
Muscle Fiber Type ◽  
Rna Seq ◽  
Type Transformation ◽  
Slow Twitch Muscle ◽  
Slow Twitch

Muscle development requires myoblast differentiation and muscle fiber formation. Myod family inhibitor (Mdfi) inhibits myogenic regulatory factors in NIH3T3 cells, but how Mdfi regulates myoblast myogenic development is still unclear. In the present study, we constructed an Mdfi-overexpression (Mdfi-OE) C2C12 cell line by the CRISPR/Cas9 system and performed RNA-seq on Mdfi-OE and wild-type (WT) C2C12 cells. The RNA-seq results showed that the calcium signaling pathway was the most significant. We also established the regulatory networks of Mdfi-OE on C2C12 cell differentiation and muscle fiber type transformation and identified hub genes. Further, both RNA-seq and experimental verification demonstrated that Mdfi promoted C2C12 cell differentiation by upregulating the expression of Myod, Myog, and Myosin. We also found that the positive regulation of Mdfi on fast-to-slow-twitch muscle fiber transformation is mediated by Myod, Camk2b, and its downstream genes, such as Pgc1a, Pdk4, Cs, Cox4, Acadm, Acox1, Cycs, and Atp5a1. In conclusion, our results demonstrated that Mdfi promotes C2C12 cell differentiation and positively modulates fast-to-slow-twitch muscle fiber transformation. These findings further our understanding of the regulatory mechanisms of Mdfi in myogenic development and muscle fiber type transformation. Our results suggest potential therapeutic targets for muscle- and metabolic-related diseases.

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