Dataset Information


Transcriptional analysis of genes regulating the mitochondrial dNTP pool in muscle cells.

ABSTRACT: Mutations in genes involved in dNTP metabolism can lead to tissue-specific mitochondrial depletion syndromes (MDS), likely because the expression of key enzymes is reduced to critical levels in post mitotic cells. Our goal was to establish an in vitro skeletal muscle cell model to study the muscle specificity of MDS associated with mitochondrial dNTP pool imbalance. We performed a comprehensive analysis at the mRNA level of enzymes and transporters responsible for dNTP pool imbalance in muscle cells in vitro and in vivo. Agilent Mouse Oligo Arrays 4x44K were utilized to examine expression levels in proliferating and differentiated C2C12 cells as well as in the mouse EDL (fast glycolytic) and soleus (slow oxidative) muscles. The comparison of mRNA expression profiles supports the reliability of our in vitro cell system. Proliferating mouse C2C12 myoblasts were collected at about 50 percent confluence. Myoblasts were then induced to differentiate in vitro into myotubes that were harvested after 96 hours and further purified to reduce the contribution of mononucleated cells present in the culture. Gene expression in C2C12 myoblasts and myotubes was compared with fully differentiated muscle fibers in vivo. To this aim, the extensor digitorum longus (EDL) and soleus hind limb muscles were isolated from adult CD1 mice. These muscles were selected because they have different metabolic (glycolytic vs. oxidative) and twitching (fast vs. slow) properties. Triplicate total RNA samples were submitted to gene expression profiling.

ORGANISM(S): Mus musculus  

SUBMITTER: Francesco Chemello   Miriam Frangini  Stefano Cagnin  Beniamina Pacchioni  Chiara Rampazzo  Paolo Laveder  Caterina Millino  Francesco Martinati  Camilla Bean  Barbara Celegato  Gerolamo Lanfranchi 

PROVIDER: E-GEOD-41877 | ArrayExpress | 2013-03-08



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Synthesis of mitochondrial DNA precursors during myogenesis, an analysis in purified C2C12 myotubes.

Frangini Miriam M   Franzolin Elisa E   Chemello Francesco F   Laveder Paolo P   Romualdi Chiara C   Bianchi Vera V   Rampazzo Chiara C  

The Journal of biological chemistry 20130107 8

During myogenesis, myoblasts fuse into multinucleated myotubes that acquire the contractile fibrils and accessory structures typical of striated skeletal muscle fibers. To support the high energy requirements of muscle contraction, myogenesis entails an increase in mitochondrial (mt) mass with stimulation of mtDNA synthesis and consumption of DNA precursors (dNTPs). Myotubes are quiescent cells and as such down-regulate dNTP production despite a high demand for dNTPs. Although myogenesis has bee  ...[more]

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