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

Scientific Reports
Arun J SinghChrissa Kioussi

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 ...Continue Reading

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Citations

May 23, 2018·Journal of Developmental Biology·Chih-Ning Chang, Chrissa Kioussi

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Datasets Mentioned

BETA
SRP126903

Methods Mentioned

BETA
fluorescent-activated cell sorting
transgenic
Fluorescent
FACS
PCA
RNA-Seq
single cell sequencing

Software Mentioned

R
pheatmap
TopHat
Cytoscape
HTseq
R software
DESeq2
Casava
Panther
Fastq

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