A Noninvasive In Vitro Monitoring System Reporting Skeletal Muscle Differentiation

The cell–cell adhesion molecule N-cadherin, with its associated catenins, is expressed by differentiating skeletal muscle and its precursors. Although N-cadherin's role in later events of skeletal myogenesis such as adhesion during myoblast fusion is well established, less is known about its role in earlier events such as commitment and differentiation. Using an in vitro model system, we have determined that N-cadherin– mediated adhesion enhances skeletal muscle differentiation in three-dimensional cell aggregates. We transfected the cadherin-negative BHK fibroblastlike cell line with N-cadherin. Expression of exogenous N-cadherin upregulated endogenous β-catenin and induced strong cell–cell adhesion. When BHK cells were cultured as three-dimensional aggregates, N-cadherin enhanced withdrawal from the cell cycle and stimulated differentiation into skeletal muscle as measured by increased expression of sarcomeric myosin and the 12/101 antigen. In contrast, N-cadherin did not stimulate differentiation of BHK cells in monolayer cultures. The effect of N-cadherin was not unique since E-cadherin also increased the level of sarcomeric myosin in BHK aggregates. However, a nonfunctional mutant N-cadherin that increased the level of β-catenin failed to promote skeletal muscle differentiation suggesting an adhesion-competent cadherin is required. Our results suggest that cadherin-mediated cell–cell interactions during embryogenesis can dramatically influence skeletal myogenesis.

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Static magnetic fields enhance skeletal muscle differentiation in vitro by improving myoblast alignment

Cytometry Part A ◽

10.1002/cyto.a.20447 ◽

2007 ◽

Vol 71A (10) ◽

pp. 846-856 ◽

Cited By ~ 41

Author(s):

Dario Coletti ◽

Laura Teodori ◽

Maria C. Albertini ◽

Marco Rocchi ◽

Alessandro Pristerà ◽

...

Keyword(s):

Skeletal Muscle ◽

Magnetic Fields ◽

Muscle Differentiation ◽

Skeletal Muscle Differentiation ◽

Static Magnetic Fields

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6-Bromoindirubin-3′-oxime intercepts GSK3 signaling to promote and enhance skeletal muscle differentiation affecting miR-206 expression in mice

Scientific Reports ◽

10.1038/s41598-019-54574-4 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 7

Author(s):

Elvira Ragozzino ◽

Mariarita Brancaccio ◽

Antonella Di Costanzo ◽

Francesco Scalabrì ◽

Gennaro Andolfi ◽

...

Keyword(s):

Skeletal Muscle ◽

Myogenic Differentiation ◽

Muscle Differentiation ◽

C2c12 Cells ◽

Skeletal Muscle Differentiation ◽

Myoblast Proliferation ◽

Enhanced Expression ◽

Pharmacologically Active

AbstractDystrophies are characterized by progressive skeletal muscle degeneration and weakness as consequence of their molecular abnormalities. Thus, new drugs for restoring skeletal muscle deterioration are critically needed. To identify new and alternative compounds with a functional role in skeletal muscle myogenesis, we screened a library of pharmacologically active compounds and selected the small molecule 6-bromoindirubin-3′-oxime (BIO) as an inhibitor of myoblast proliferation. Using C2C12 cells, we examined BIO’s effect during myoblast proliferation and differentiation showing that BIO treatment promotes transition from cell proliferation to myogenic differentiation through the arrest of cell cycle. Here, we show that BIO is able to promote myogenic differentiation in damaged myotubes in-vitro by enriching the population of newly formed skeletal muscle myotubes. Moreover, in-vivo experiments in CTX-damaged TA muscle confirmed the pro-differentiation capability of BIO as shown by the increasing of the percentage of myofibers with centralized nuclei as well as by the increasing of myofibers number. Additionally, we have identified a strong correlation of miR-206 with BIO treatment both in-vitro and in-vivo: the enhanced expression of miR-206 was observed in-vitro in BIO-treated proliferating myoblasts, miR-206 restored expression was observed in a forced miR-206 silencing conditions antagomiR-mediated upon BIO treatment, and in-vivo in CTX-injured muscles miR-206 enhanced expression was observed upon BIO treatment. Taken together, our results highlight the capacity of BIO to act as a positive modulator of skeletal muscle differentiation in-vitro and in-vivo opening up a new perspective for novel therapeutic targets to correct skeletal muscle defects.

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Remodelling of the nuclear lamina and nucleoskeleton is required for skeletal muscle differentiation in vitro

Journal of Cell Science ◽

10.1242/jcs.01630 ◽

2005 ◽

Vol 118 (2) ◽

pp. 409-420 ◽

Cited By ~ 70

Author(s):

E. Markiewicz

Keyword(s):

Skeletal Muscle ◽

Nuclear Lamina ◽

Muscle Differentiation ◽

Skeletal Muscle Differentiation

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The SMYD3 methyltransferase promotes myogenesis by activating the myogenin regulatory network

Scientific Reports ◽

10.1038/s41598-019-53577-5 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 4

Author(s):

Roberta Codato ◽

Martine Perichon ◽

Arnaud Divol ◽

Ella Fung ◽

Athanassia Sotiropoulos ◽

...

Keyword(s):

Skeletal Muscle ◽

Muscle Development ◽

Myogenic Differentiation ◽

Muscle Differentiation ◽

Skeletal Muscle Differentiation ◽

Skeletal Muscle Development ◽

Genome Wide ◽

Coordinated Expression

AbstractThe coordinated expression of myogenic regulatory factors, including MyoD and myogenin, orchestrates the steps of skeletal muscle development, from myoblast proliferation and cell-cycle exit, to myoblast fusion and myotubes maturation. Yet, it remains unclear how key transcription factors and epigenetic enzymes cooperate to guide myogenic differentiation. Proteins of the SMYD (SET and MYND domain-containing) methyltransferase family participate in cardiac and skeletal myogenesis during development in zebrafish, Drosophila and mice. Here, we show that the mammalian SMYD3 methyltransferase coordinates skeletal muscle differentiation in vitro. Overexpression of SMYD3 in myoblasts promoted muscle differentiation and myoblasts fusion. Conversely, silencing of endogenous SMYD3 or its pharmacological inhibition impaired muscle differentiation. Genome-wide transcriptomic analysis of murine myoblasts, with silenced or overexpressed SMYD3, revealed that SMYD3 impacts skeletal muscle differentiation by targeting the key muscle regulatory factor myogenin. The role of SMYD3 in the regulation of skeletal muscle differentiation and myotube formation, partially via the myogenin transcriptional network, highlights the importance of methyltransferases in mammalian myogenesis.

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Inositide-Dependent Phospholipase C Signaling Mimics Insulin in Skeletal Muscle Differentiation by Affecting Specific Regions of the Cyclin D3 Promoter

Endocrinology ◽

10.1210/en.2006-1003 ◽

2007 ◽

Vol 148 (3) ◽

pp. 1108-1117 ◽

Cited By ~ 42

Author(s):

Irene Faenza ◽

Giulia Ramazzotti ◽

Alberto Bavelloni ◽

Roberta Fiume ◽

Gian Carlo Gaboardi ◽

...

Keyword(s):

Skeletal Muscle ◽

Phospholipase C ◽

Myogenic Differentiation ◽

Muscle Differentiation ◽

C2c12 Cells ◽

Cyclin D3 ◽

Expression Vectors ◽

Skeletal Muscle Differentiation ◽

Small Interfering Rnas

Our main goal in this study was to investigate the role of phospholipase C (PLC) β1 and PLCγ1 in skeletal muscle differentiation and the existence of potential downstream targets of their signaling activity. To examine whether PLC signaling can modulate the expression of cyclin D3, a target of PLCβ1 in erythroleukemia cells, we transfected C2C12 cells with expression vectors containing PLCβ1 or PLCγ1 cDNA and with small interfering RNAs from regions of the PLCβ1 or PLCγ1 gene and followed myogenic differentiation in this well-established cell system. Intriguingly, overexpressed PLCβ1 and PLCγ1 were able to mimic insulin induction of both cyclin D3 and muscle differentiation. By knocking down PLCβ1 or PLCγ1 expression, C2C12 cells almost completely lost the increase in cyclin D3, and the differentiation program was down-regulated. To explore the induction of the cyclin D3 gene promoter during this process, we used a series of 5′-deletions of the 1.68-kb promoter linked to a reporter gene and noted a 5-fold augmentation of promoter activity upon insulin stimulation. These constructs were also cotransfected with PLCβ1 or PLCγ1 cDNAs and small interfering RNAs, respectively. Our data indicate that PLCβ1 or PLCγ1 signaling is capable of acting like insulin in regard to both the myogenic differentiation program and cyclin D3 up-regulation. Taken together, this is the first study that hints at cyclin D3 as a target of PLCβ1 and PLCγ1 during myogenic differentiation in vitro and implies that up-regulation of these enzymes is sufficient to mimic the actions of insulin in this process.

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Titin expression as an early indication of heart and skeletal muscle differentiation in vitro. Developmental re-organisation in relation to cytoskeletal constituents

Journal of Muscle Research and Cell Motility ◽

10.1007/bf00140321 ◽

1996 ◽

Vol 17 (1) ◽

pp. 23-36 ◽

Cited By ~ 13

Author(s):

Frank T. L. Van der Loop ◽

Guillaume J. J. M. van Eys ◽

Gert Schaart ◽

Frans C. S. Ramaekers

Keyword(s):

Skeletal Muscle ◽

Muscle Differentiation ◽

Skeletal Muscle Differentiation ◽

Early Indication

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TGF-β Inhibits Muscle Differentiation by Blocking Autocrine Signaling Pathways Initiated by IGF-II

Molecular Endocrinology ◽

10.1210/me.2010-0292 ◽

2011 ◽

Vol 25 (1) ◽

pp. 128-137 ◽

Cited By ~ 25

Author(s):

Samantha Gardner ◽

Damir Alzhanov ◽

Paul Knollman ◽

David Kuninger ◽

Peter Rotwein

Keyword(s):

Skeletal Muscle ◽

Signaling Pathways ◽

Muscle Differentiation ◽

Receptor Activation ◽

Skeletal Muscle Differentiation ◽

Myotube Formation ◽

Gene And Protein Expression ◽

Igf I ◽

Tgf Β1

Skeletal muscle differentiation and regeneration are regulated by interactions between exogenous hormone- and growth factor-activated signaling cascades and endogenous muscle-specific transcriptional programs. IGF-I and IGF-II can promote muscle differentiation in vitro and can enhance muscle maintenance and repair in vivo. In contrast, members of the TGF-β superfamily prominently inhibit muscle differentiation and regeneration. In this study, we have evaluated functional interactions between IGF- and TGF-β-regulated signaling pathways during skeletal muscle differentiation. In the mouse C2 muscle cell line and in human myoblasts in primary culture, addition of TGF-β1 blocked differentiation in a dose-dependent way, inhibited expression of muscle-specific mRNAs and proteins, and impaired myotube formation. TGF-β1 also diminished stimulation of IGF-II gene expression in myoblasts, decreased IGF-II secretion, and reduced IGF-I receptor activation. To test the hypothesis that TGF-β1 prevents muscle differentiation primarily by blocking IGF-II production, we examined effects of IGF analogues on TGF-β actions in myoblasts. Although both IGF-I and IGF-II restored muscle gene and protein expression, and stimulated myotube formation in the presence of TGF-β1, they did not reduce TGF-β1-stimulated signaling, as measured by no decline in phosphorylation of SMA and mothers against decapentaplegic homolog (Smad)3, or in induction of TGF-β-activated target genes, including a Smad-dependent promoter-reporter plasmid. Our results demonstrate that TGF-β disrupts an IGF-II-stimulated autocrine amplification cascade that is necessary for muscle differentiation in vitro. Because this inhibitory pathway can be overcome by exogenous IGFs, our observations point toward potential strategies to counteract disorders that reduce muscle mass and strength.

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