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. 2021 Jul 7:12:643497.
doi: 10.3389/fgene.2021.643497. eCollection 2021.

The Expression Profiles of mRNAs and lncRNAs in Buffalo Muscle Stem Cells Driving Myogenic Differentiation

Ruimen Zhang  1 , Jinling Wang  1 , Zhengzhong Xiao  2 , Chaoxia Zou  1 , Qiang An  1 , Hui Li  1 , Xiaoqing Zhou  2 , Zhuyue Wu  2 , Deshun Shi  1 , Yanfei Deng  1 , Sufang Yang  1   3 , Yingming Wei  1
Affiliations

The Expression Profiles of mRNAs and lncRNAs in Buffalo Muscle Stem Cells Driving Myogenic Differentiation

Ruimen Zhang et al. Front Genet. .

Abstract

Buffalo breeding has become an important branch of the beef cattle industry. Hence, it is of great significance to study buffalo meat production and meat quality. However, the expression profiles of mRNA and long non-coding RNAs (lncRNA) molecules in muscle stem cells (MuSCs) development in buffalo have not been explored fully. We, therefore, performed mRNA and lncRNA expression profiling analysis during the proliferation and differentiation phases of MuSCs in buffalo. The results showed that there were 4,820 differentially expressed genes as well as 12,227 mRNAs and 1,352 lncRNAs. These genes were shown to be enriched in essential biological processes such as cell cycle, p53 signaling pathway, RNA transport and calcium signaling pathway. We also identified a number of functionally important genes, such as MCMC4, SERDINE1, ISLR, LOC102394806, and LOC102403551, and found that interference with MYLPF expression significantly inhibited the differentiation of MuSCs. In conclusion, our research revealed the characteristics of mRNA and lncRNA expression during the differentiation of buffalo MuSCs. This study can be used as an important reference for the study of RNA regulation during muscle development in buffalo.

Keywords: buffalo; mRNAs; muscle stem cells; myogenesis; non-coding RNAs.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Analysis of the characteristics of buffalo MuSCs during proliferation and differentiation. (A) Proliferation phenotype of muscle stem cells (GM samples). (B) Differentiation phenotype of muscle stem cells (DM samples). (C) MuSCs grown as GM and DM samples were subjected to real-time PCR for BCL-2 and Pax7. (D) The differentiation phenotype (DM) samples of MuSCs were subjected to real-time PCR for MyoD1, MyoG, and MyHC. (E,F) MuSCs grown as GM and DM samples were subjected to western blotting for determination of PCNA and CDK2 proteins. (G) The differentiation phenotype (DM) samples of MuSCs were subjected to immunofluorescence for MyOD1. The data are presented as means ± SDs, n = 3 per group. *P < 0.05; **P < 0.01; ***P < 0.001. Scale bars = 100/200 μm.
Figure 2
Figure 2
RNA-seq analysis of buffalo MuSCs during proliferation and differentiation. (A) RNA-seq information statistics of samples. (B) Mapped reads distribution. (C) The distribution for the reads in different chromosomes.
Figure 3
Figure 3
Gene ontology (GO) analysis. (A) GO analysis of DEGs in the GM samples of MuSCs. (B) GO analysis of DEGs in the DM samples of MuSCs.
Figure 4
Figure 4
Kyoto Encyclopedia of Genes and Genomes pathway analysis. (A) Top 30 KEGG terms of DEGs in the GM samples of MuSCs. (B) Top 30 KEGG terms of DEGs in the DM samples of MuSCs.
Figure 5
Figure 5
Validation of the expression levels of DEGs, DE lncRNAs by RT-qPCR. (A,B) The validation results of DEGs and (C) DE lncRNAs. The data are present as means ± SDs, n = 3 per group. ***P < 0.001.
Figure 6
Figure 6
Expression and characterization of MYLPF in buffalo skeletal muscle. (A) Validation of the expression of MYLPF by RT-qPCR. (B) The expression levels of MYLPF in different tissues of buffalo. (C) The interference efficiency of MYLPF was measured by RT-qPCR. (D) Inhibition of MYLPF expression on myotubule formation. (E) The mRNA expression of myogenesis marker gene, MyOD1, was measured by RT-qPCR. The data are presented as means ± SDs, n = 3 per group. *P < 0.05; **P < 0.01. Scale bars = 100 μm.
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References

    1. Bass J. J., Nakhuda A., Deane C. S., Brook M. S., Wilkinson D. J., Phillips B. E., et al. . (2020). Overexpression of the vitamin D receptor (VDR) induces skeletal muscle hypertrophy. Mol. Metab. 42:101059. 10.1016/j.molmet.2020.101059 - DOI - PMC - PubMed
    1. Bhat Z. F., Kumar S., Bhat H. F. (2017). In vitro meat: a future animal-free harvest. Crit. Rev. Food Sci. Nutr. 57, 782–789. 10.1080/10408398.2014.924899 - DOI - PubMed
    1. Bonny S. P. F., Gardner G. E., Pethick D. W., Hocquette J.-F. (2015). What is artificial meat and what does it mean for the future of the meat industry? J. Integr. Agric. 14, 255–263. 10.1016/S2095-3119(14)60888-1 - DOI
    1. Boscolo Sesillo F., Wong M., Cortez A., Alperin M. (2020). Isolation of muscle stem cells from rat skeletal muscles. Stem. Cell Res. 43:101684. 10.1016/j.scr.2019.101684 - DOI - PMC - PubMed
    1. Chen R., Lei S., Jiang T., Zeng J., Zhou S., She Y. (2020). Roles of lncRNAs and circRNAs in regulating skeletal muscle development. Acta Physiol. 228:e13356. 10.1111/apha.13356 - DOI - PubMed