FACS-Seq analysis of Pax3-derived cells identifies non-myogenic lineages in the embryonic forelimb

Sci Rep. 2018 May 16;8(1):7670. doi: 10.1038/s41598-018-25998-1.

Abstract

Skeletal muscle in the forelimb develops during embryonic and fetal development and perinatally. While much is known regarding the molecules involved in forelimb myogenesis, little is known about the specific mechanisms and interactions. Migrating skeletal muscle precursor cells express Pax3 as they migrate into the forelimb from the dermomyotome. To compare gene expression profiles of the same cell population over time, we isolated lineage-traced Pax3+ cells (Pax3 EGFP ) from forelimbs at different embryonic days. We performed whole transcriptome profiling via RNA-Seq of Pax3+ cells to construct gene networks involved in different stages of embryonic and fetal development. With this, we identified genes involved in the skeletal, muscular, vascular, nervous and immune systems. Expression of genes related to the immune, skeletal and vascular systems showed prominent increases over time, suggesting a non-skeletal myogenic context of Pax3-derived cells. Using co-expression analysis, we observed an immune-related gene subnetwork active during fetal myogenesis, further implying that Pax3-derived cells are not a strictly myogenic lineage, and are involved in patterning and three-dimensional formation of the forelimb through multiple systems.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation
  • Cell Lineage / genetics*
  • Cells, Cultured
  • Embryo, Mammalian / cytology*
  • Embryo, Mammalian / metabolism
  • Female
  • Forelimb / embryology*
  • Forelimb / metabolism
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental*
  • Gene Regulatory Networks
  • Mice
  • Mice, Inbred ICR
  • Muscle Development / genetics
  • Muscle Proteins / genetics*
  • Muscle Proteins / metabolism
  • Muscle, Skeletal / cytology*
  • Muscle, Skeletal / metabolism
  • Myoblasts / cytology
  • Myoblasts / metabolism
  • PAX3 Transcription Factor / genetics
  • PAX3 Transcription Factor / metabolism*

Substances

  • Muscle Proteins
  • PAX3 Transcription Factor
  • Pax3 protein, mouse