The Mechanism of Effect of Hypoxia on Myofiber Type Transformation in Chicken Myoblasts
The cobalt chloride(CoCl2)was utilized in this study to simulate hypoxia in muscle cells and employed transcriptomic sequencing to analyze the effects of hypoxia on muscle fiber type and genes expression that related to muscle fiber types.Primary chicken myoblasts were treated with different concentrations of CoCl2(0,50,100,200 and 400 μmol·L-1),with 3 repeat wells in each group.The CoCl2 concentration was screened by detecting the expression of hypoxia marker genes and observing cell morphology to determine the most suitable concentration for modeling.Subsequently,the collected cell was used to detect the gene expression profile of chicken myoblast in hypoxia treatment and control treatment by Illumina technology and assess differential gene expression and enriched signaling pathways between treatments.The results in-dicated that 200 μmol·L-1 CoCl2 was the most suitable concentration for constructing the hypoxic model in chicken embryonic myogenic cells.Transcriptomic results showed a total of 1 474 and 2 028 differentially expressed genes in the hypoxia treatment compared to the control group at 24 and 48 h,respectively.Interestingly,959 genes showed differential expression in both time points.GO and KEGG analyses revealed significant alterations in key factors related to signaling pathways such as regulation of signaling receptor activity,integral component of plasma mem-brane,neuroactive ligand-receptor interaction,PI3K-AKT signaling pathway and muscle contrac-tion in chicken muscle cells under hypoxic conditions.Consistent findings from sequencing and real-time quantitative polymerase chain reaction(qRT-PCR)demonstrated that slow muscle fiber-specific genes such as MYH7B and CSRP2 were significantly upregulated after hypoxia treatment,while fast muscle fiber-specific genes including MYH1F,MYH1D,TNNI2,and SOX6 were significantly downregulated,which indicated a transition of chicken muscle cells from fast to slow muscle fibers was induced by hypoxia.This study provides preliminary insights into the mechanisms underlying the hypoxia-induced transition of muscle fiber types in chicken myo-genic cells and offers a valuable gene expression database for future research on poultry meat quality and hypoxia-related studies in mammals.
hypoxiachicken myoblastsmuscle fiber type transformation
万方数据
维普
