缺氧对鸡成肌细胞肌纤维类型转化作用的机制探究

doi:10.11843/j.issn.0366-6964.2024.06.012

Abstract

Abstract:

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 indicated 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 membrane, neuroactive ligand-receptor interaction, PI3K-AKT signaling pathway and muscle contraction 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 myogenic cells and offers a valuable gene expression database for future research on poultry meat quality and hypoxia-related studies in mammals.

Key words: hypoxia, chicken myoblasts, muscle fiber type transformation

CLC Number:

S831

Binghong XIE, Yifan LIU, Fuguang XUE, Yanju SHAN, Yunjie TU, Gaige JI, Xiaojun JU, Jingting SHU, Hongxiang WU. The Mechanism of Effect of Hypoxia on Myofiber Type Transformation in Chicken Myoblasts[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(6): 2397-2408.

Figures/Tables 7

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基因 Gene	引物序列(5′→3′) Primer sequence	产物长度/bp Product length	退火温度/℃ Tm
MYH1F	F：CTTGTGGATGGTTGTTCGCA； R：CCTCCTGTTCCAGCACAAAC	191	56
MYH7B	F：CAGCAGCCATTCCAGATGAC； R：TCTTGGCCAGACGTTCATCT	171	52
HIF1A	F：CGTGTAAAGGCGTGCAAAAC； R：CTGTTGCCTTGTATGGGAGC	170	60
VEGFA	F：AGTCTACGAACGCAGCTTCT； R：ACAGGGACACATTCTAGGCC	156	60
CSRP2	F：ATGGACAGGGGAGAGAGACT； R：TTTCTCCGCCGCATAAACAG	153	60
CSRP3	F：CCTCCACACCAACTAACCCT； R：TTCCCACAAATAGCACAGCG	154	60
MYBPC2	F：CCTCATCATCAACGAAGCGG； R：TGGATCCTCTTGCTGGGTTT	240	60
MYH1D	F：CACTTCTCCCTGGTGCACTA； R：AGACCCCTTCTTCTTGCCTC	207	60
MYH1E	F：AGCAAGTGGACGATCTGGAA； R：TTTGCTCTGGATCTGGCTGA	194	60
PRKAG3	F：CACAAGCGCATCCTCAAGTT； R：TGACCGGCAGCATTAACAAC	203	60
SOX6	F：GCTTTCCCTGACATGCACAA； R：AGGTACGTTTTGGTCGAGGT	169	60
TNNI2	F：GCAAAACTACCTGGCAGAGC； R：CCCTCAGGTCAAACAGCTTC	191	60
VEGFC	F：CGAACAAAACGCTGTGATGC； R：TTGTCTCTGAAAGCCCCACA	214	60
MYOG	F：GAGGGGCTTTGGAGGAGAAG； R：TTCACCTTCTTCAGCCTCCG	154	60
PGM1	F：AGTTTTGGTACTGGCTCCGA； R：CACAAAGGAGCGGTCAAACA	234	60
PPARGC1A	F：AAAGACGTCCCTGCTCTGAA； R：TGGCTTGCAGGTGATTTGTC	153	60

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The Mechanism of Effect of Hypoxia on Myofiber Type Transformation in Chicken Myoblasts

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