Purinergic Signalling

, Volume 8, Issue 2, pp 207–221 | Cite as

Transcriptional profile of GTP-mediated differentiation of C2C12 skeletal muscle cells

  • Rosa Mancinelli
  • Tiziana Pietrangelo
  • Geoffrey Burnstock
  • Giorgio Fanò
  • Stefania Fulle
Original Article

Abstract

Several purine receptors have been localised on skeletal muscle membranes. Previous data support the hypothesis that extracellular guanosine 5′-triphosphate (GTP) is an important regulatory factor in the development and function of muscle tissue. We have previously described specific extracellular binding sites for GTP on the plasma membrane of mouse skeletal muscle (C2C12) cells. Extracellular GTP induces an increase in intracellular Ca2+ concentrations that results in membrane hyperpolarisation through Ca2+-activated K+ channels, as has been demonstrated by patch-clamp experiments. This GTP-evoked increase in intracellular Ca2+ is due to release of Ca2+ from intracellular inositol-1,4,5-trisphosphate-sensitive stores. This enhances the expression of the myosin heavy chain in these C2C12 myoblasts and commits them to fuse into multinucleated myotubes, probably via a phosphoinositide-3-kinase-dependent signal-transduction mechanism. To define the signalling of extracellular GTP as an enhancer or modulator of myogenesis, we investigated whether the gene-expression profile of differentiated C2C12 cells (4 and 24 h in culture) is affected by extracellular GTP. To investigate the nuclear activity and target genes modulated by GTP, transcriptional profile analysis and real-time PCR were used. We demonstrate that in the early stages of differentiation, GTP up-regulates genes involved in different pathways associated with myogenic processes, including cytoskeleton structure, the respiratory chain, myogenesis, chromatin reorganisation, cell adhesion, and the Jak/Stat pathway, and down-regulates the mitogen-activated protein kinase pathway. GTP also increases the expression of three genes involved in myogenesis, Pp3ca, Gsk3b, and Pax7. Our data suggests that in the myogenic C2C12 cell line, extracellular GTP acts as a differentiative factor in the induction and sustaining of myogenesis.

Keywords

Transcriptome Myogenesis GTP 

Abbreviations

BSA

Bovine serum albumin

[Ca2+]i

Intracellular calcium ion concentration

DM

Standard differentiating medium

GTP

Guanosine 5′-triphosphate

MRFs

Myogenic regulatory factors

MyHC

Myosin heavy chain

SM

Synthetic differentiating medium

RB2

Reactive blue 2

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Rosa Mancinelli
    • 1
    • 2
    • 3
  • Tiziana Pietrangelo
    • 1
    • 2
    • 4
  • Geoffrey Burnstock
    • 5
  • Giorgio Fanò
    • 1
    • 2
    • 4
  • Stefania Fulle
    • 1
    • 2
    • 3
  1. 1.Department of Neuroscience and ImagingUniversity “G. d’Annunzio” Chieti-PescaraChietiItaly
  2. 2.Interuniversity Institute of Myology (IIM)ChietiItaly
  3. 3.Cellular Physiology Unit, Center for Excellence on Ageing (CeSI)“G. d’Annunzio” FoundationChietiItaly
  4. 4.Human Physiology Unit, Center for Excellence on Ageing (CeSI),-Clinical Research Center (CRC)“G. d’Annunzio” FoundationChietiItaly
  5. 5.Autonomic Neuroscience CentreUniversity College Medical SchoolLondonUK

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