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Transcriptional profiles of skeletal muscles in mice exposed to acute hypoxia

ABSTRACT: To understand the underlying gene changes in in vivo muscles during acute hypoxic exposure, we studied the transcriptional profiles of in vivo skeletal muscles from mice exposed to 8% O2 for 0 hours, 2 hours, and 6 hours. For each condition, we extracted total RNA from fresh in vivo plantaris muscles for 3 biological replicates. The NGS data was acquired using paired-end 100bp by Illumina HiSeq 2000 with a depth around 100X. The quality control of raw data indicated excellent quality of sequencing data with quality scores in the range of 33-40 all over the reads. The analysis of differential expressed genes indicated quick genes responses in skeletal muscles in both 2 and 6 hours of hypoxia. The gene changes indicated a shift from genes associated with extracellular lumen in 2 hours of hypoxia to genes associated with the nuclear lumen in 6 hours of hypoxia. Signaling pathways such as PI3K/Akt signaling, mTOR signaling, MAPK signaling showed enriched gene responses. The KEGG-pathway based pathway-network analysis suggested the PI3K/Akt signaling pathway as a nodal pathway among the gene response-enriched pathways during acute hypoxia. Our Western blot experiments indicated a functional down-regulation of PI3K/Akt signaling by de-phosphorylation of Akt.

ORGANISM(S): Mus musculus

PROVIDER: GSE81286 | GEO | 2017/10/23

SECONDARY ACCESSION(S): PRJNA321188

REPOSITORIES: GEO

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Project description:When skin tissue is damaged, the wound microenvironment is hypoxic or anoxic due to the rupture of blood vessels and the high oxygen consumption of related cells; so far, it is not clear whether such hypoxic microenvironment will promote epidermal cell migration and the underlying molecular regulatory mechanisms of this effect remains unclear. Studies to date have shown that, immortal keratinocyte cell line HaCaT and primary human keratinocytes are maintained under conditions of hypoxia (1% oxygen) or normoxia. The researchers would adopt methods such as live cell imaging system, Western blotting, transwell assays and wound scratch assays etc. to study the changes of cell migration. Expression profile of mRNAs in HaCaT cells from 3 hypoxia and 3 normoxic conditions were analyzed by the Clariom D microarray assay. Gene Ontology (GO) and KEGG genomics analyses were performed to identify significant functions, pathways, and the associations of differentially expressed mRNAs. Moreover, the potential mechanism is discussed and studied. According to relevant results, the epidermal cell migration is promoted in the early hypoxia. Furthermore, experiment showed that 1456 mRNAs showed differential expression between the hypoxia and normoxic conditions, which included 537 upregulated and 919 downregulated mRNAs. These results also shown that all G proteins are upregulated. At the same time, GO analysis indicated that most upregulated mRNA are in connection with cell inflammation and migration. Pathway analysis indicated that 20 pathways corresponded to the upregulated mRNA and 6 pathways corresponded to the upregulated mRNA. These pathway analyses have indicated that genes involved in inflammation, migration and PI3K-Akt signaling are potentially regulated. Combine with microarray assay, we further shown that G protein accelerates epidermal cell migration via activation of PI3K/Akt-dependent mTORC1 signaling under the condition of hypoxia. Based on this evidence, researchers can further reveal the molecular and cellular mechanisms of local wound hypoxia, and for improving the wound healing.

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Transcriptional profiles of skeletal muscles in mice exposed to acute hypoxia

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