Transcriptomic analysis of mouse limb tendon cells during development

Development. 2014 Oct;141(19):3683-96. doi: 10.1242/dev.108654. Epub 2014 Sep 5.

Abstract

The molecular signals driving tendon development are not fully identified. We have undertaken a transcriptome analysis of mouse limb tendon cells that were isolated at different stages of development based on scleraxis (Scx) expression. Microarray comparisons allowed us to establish a list of genes regulated in tendon cells during mouse limb development. Bioinformatics analysis of the tendon transcriptome showed that the two most strongly modified signalling pathways were TGF-β and MAPK. TGF-β/SMAD2/3 gain- and loss-of-function experiments in mouse limb explants and mesenchymal stem cells showed that TGF-β signalling was sufficient and required via SMAD2/3 to drive mouse mesodermal stem cells towards the tendon lineage ex vivo and in vitro. TGF-β was also sufficient for tendon gene expression in late limb explants during tendon differentiation. FGF does not have a tenogenic effect and the inhibition of the ERK MAPK signalling pathway was sufficient to activate Scx in mouse limb mesodermal progenitors and mesenchymal stem cells.

Keywords: ERK; Limb; Mouse; SMAD2/3; Scleraxis; TGF-β; Tendon; Transcriptome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Computational Biology
  • Extremities / physiology*
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental / genetics
  • Gene Expression Regulation, Developmental / physiology*
  • In Situ Hybridization
  • Mesenchymal Stem Cells / metabolism
  • Mice
  • Microarray Analysis
  • Mitogen-Activated Protein Kinases / metabolism
  • Real-Time Polymerase Chain Reaction
  • Signal Transduction / physiology*
  • Tendons / cytology*
  • Tendons / metabolism
  • Transcriptome / genetics
  • Transcriptome / physiology*
  • Transforming Growth Factor beta / metabolism

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • Scx protein, mouse
  • Transforming Growth Factor beta
  • Mitogen-Activated Protein Kinases