过表达MyBPC1对牛骨骼肌卫星细胞增殖分化的影响

doi:10.11843/j.issn.0366-6964.2021.012.011

Abstract

Abstract: The study aimed to explore the effects of myosin binding protein C1 (MyBPC1) on the proliferation and myogenic differentiation of bovine skeletal muscle satellite cells, and provide a basis for further study of the regulatory role of MyBPC1 in cell differentiation and muscle development. The Simmental fetal bovine primary bovine skeletal muscle satellite cells to inducing myogenic differentiation model in vitro was used to simulate the growth and development process of bovine skeletal muscle. qRT-PCR and Western blot were used to detect the cellular temporal expression profile of MyBPC1. There were two groups in this experiment. At the RNA level, there were 4 replicates in each group, and 20 μL in each replicate. However, at the protein level, there were 3 replicates in each group, and 15 μg in each replicate. qRT-PCR and Western blot were used to detect the overexpression effect of MyBPC1 transfected into bovine skeletal muscle sate-llite cells, and to further detect the expression changes of cell proliferation marker factors Pax7, Ki67 and cell differentiation marker factors MyHC, MyOG. The myotube formation status of bovine skeletal muscle satellite cells was observed. The expression level of MyBPC1 was significantly different before and after the differentiation of bovine skeletal muscle satellite cells. After the differentiation of bovine skeletal muscle satellite cells, the mRNA and protein expression of MyBPC1 were significantly higher than those in the proliferation phase (P<0.01). After overexpression of MyBPC1, the number of myotubes formed by cell differentiation was significantly greater than that of the control group. There was no significant difference at the mRNA and protein expression levels of the proliferation marker factor Pax7. The mRNA and protein expression levels of the differentiation marker factor MyHC were significantly higher than that of the control group (P<0.01). Overexpression of MyBPC1 can promote the myogenic differentiation of bovine skele-tal muscle satellite cells in vitro, which lays the foundation for further studying the regulatory mechanism of MyBPC1 on bovine skeletal muscle satellite cells.

Key words: bovine, skeletal muscle satellite cells, overexpression, MyBPC1, proliferation, differentiation

CLC Number:

S823.2

HONG Qianqian, GUO Hong, GAO Shuxin, GUO Yiwen. Effect of Overexpression of MyBPC1 on Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3439-3448.

References 39

[1]	郑莉芳, 陈佩杰, 肖卫华.骨骼肌质量控制信号通路[J].生理学报, 2019, 71(4):671-679.ZHENG L F, CHEN P J, XIAO W H. Signaling pathways controlling skeletal muscle mass[J]. Sheng Li Xue Bao, 2019, 71(4):671-679.(in Chinese)
[2]	CHARGE S B P, RUDNICKI M A.Cellular and molecular regulation of muscle regeneration[J]. Physiol Rev, 2004, 84(1):209-238.
[3]	YIN H, PRICE F, RUDNICKI M A. Satellite cells and the muscle stem cell niche[J]. Physiol Rev, 2013, 93(1):23-67.
[4]	SAID R S, MUSTAFA A G, ASFOUR H A, et al.Myogenic satellite cells:Biological milieu and possible clinical applications[J].Pak J Biol Sci, 2017, 20(1):1-11
[5]	WU X, ZHU Z, YERLE M, et al. Radiation hybrid mapping of four genes (MYBPC1, LUM, ZRF1 and ATP2B4) expressed in embryo skeleton muscle to pig chromosomes 5 and 9[J]. Anim Genet, 2004, 35(6):472-473.
[6]	VERARDO L L, NASCIMENTO C S, SILVE F F, et al. Identification and validation of differentially expressed genes from pig skeletal muscle[J].J Anim Breed Genet, 2013, 130(5):372-381.
[7]	TONG B, XING Y P, MURAMATSU Y, et al. Association of expression levels in skeletal muscle and a SNP in the MYBPC1 gene with growth-related trait in Japanese Black beef cattle[J].J Genet, 2015, 94(1):135-137.
[8]	TONG B, SASAKI S, MURAMATSU Y, et al.Association of a single-nucleotide polymorphism in myosin-binding protein C, slow-type (MYBPC1) gene with marbling in Japanese Black beef cattle[J]. J Genet, 2014, 45(4):611-612.
[9]	HA K, BUCHAN J G, ALVARADO D M, et al.MYBPC1 mutations impair skeletal muscle function in zebrafish models of arthrogryposis[J].Hum Mol Genet, 2013, 22(24):4967-4977.
[10]	CHEN Z, ZHAO T J, LI J, et al.Slow skeletal muscle myosin-binding protein-C (MyBPC1) mediates recruitment of muscle-type creatine kinase (CK) to myosin[J].Biochem J, 2011, 436(2):437-445.
[11]	SEALE P, SABOURIN L A, GIRGIS-GABARDO A, et al.Pax7 is required for the specification of myogenic satellite cells[J].Cell, 2000, 102(6):777-786.
[12]	SCHMIDT M, SCHVLER S C, HVTTNE S S, et al.Adult stem cells at work:regenerating skeletal muscle[J].Cellr Mol Life Sci, 2019, 76(13):2559-2570.
[13]	MILLER I, MIN M, YANG C, et al.Ki67 is a graded rather than a binary marker of proliferation versus quiescence[J].Cell Rep, 2018, 24(5):1105-1112.
[14]	SUN X M, KAUFMAN P D. Ki-67:more than a proliferation marker[J].Chromosoma, 2018, 127(2):175-186.
[15]	MENON S S, GURUVAYOORAPPAN G, MURUGESAN K. Ki-67 protein as a tumour proliferation marker[J].Clin Chim Acta, 2019, 491:39-45.
[16]	WANG Y X, RUDNICKI M A. Satellite cells, the engines of muscle repair.[J].Nat Rev Mol Cell Biol, 2011, 13(2):127-133.
[17]	LILJA K C, ZHANG N, MAGLI A, at el.P Pax7 remodels the chromatin landscape in skeletal muscle stem cells.[J].PLoS One, 2017, 12(4):e0176190.
[18]	ZAMMIT P S. Function of the myogenic regulatory factors Myf5, MyoD, Myogenin and MRF4 in skeletal muscle, satellite cells and regenerative myogenesis[J].Semin Cell Dev Biol, 2017, 72:19-32.
[19]	HERNANDEZ J M, GONZALEZ E G, BRUNC E, at el.The myogenic regulatory factors, determinants of muscle development, cell identity and regeneration[J].Semin Cell Dev Biol, 2017, 72:10-18.
[20]	SCHIAFFINO S. Muscle fiber type diversity revealed by anti-myosin heavy chain antibodies[J].FEBS J, 2018, 285(20):3688-3694.
[21]	CHENG Q H, LI J Y, SHENG X L, et al.Higher frequency electrical stimulation enhanced myloglossus satellite cell differentiation by upregulating expression of Pax7 mRNA, MyoD, myogenin and MyHC protein[J].Eur Rev Medi Pharmacol Sci, 2020, 24(23):12041-12049.
[22]	王男, 李剑锋, 王莉晓, 等.Pax7、MyoD及MyoG基因用于推断损伤时间的初步研究[J].中国法医学杂志, 2019, 34(4):330-335.WANG N, LI J F, WANG L X, et al.Preliminary study of the effects of Pax7, MyoD and MyoG genes on predicting the dating of wound[J].Chinese Journal of Forensic Medicine, 2019, 34(4):330-335.(in Chinese)
[23]	ZHANG H F, WEN J F, BIGOT A, et al.Human myotube formation is determined by MyoD-Myomixer/Myomaker axis[J].Sci Adv, 2020, 6(51):eabc4062.
[24]	AGARWAL M, SHARMA A, KUMAR P, et al.Myosin heavy chain-embryonic regulates skeletal muscle differentiation during mammalian development[J].Development, 2020, 147(7):dev184507.
[25]	王轶敏, 代阳, 刘新峰, 等.牛骨骼肌细胞的分离鉴定和诱导分化[J].中国畜牧兽医, 2014, 41(7):142-147.WANG Y M, DAI Y, LIU X F, et al.Isolation, identification and induced differentiation of bovine skeletal muscle satellite cells[J].China Animal Husbandry & Veterinary Medicine, 2014, 41(7):142-147.(in Chinese)
[26]	UCHIDA Y, CHIBA T, RYOTA K, et al.Post-transcriptional regulation of inflammation by RNA-binding proteins via cis-elements of mRNAs[J].J Biochem, 2019, 166(5):375-382.
[27]	MOOTHA V K, BUNKENBORG J, OLSEN J V, et al.Integrated analysis of protein composition, tissue diversity, and gene regulation in mouse mitochondria[J].Cell, 2003, 115(5):629-640.
[28]	GUPTA M K, ROBBINS J.Post-translational control of cardiac hemodynamics through myosin binding protein C[J].Pflugers Arch, 2014, 466(2):231-236.
[29]	MCNAMARA J W, SADAYAPPAN S.Skeletal myosin binding protein-C:An increasingly important regulator of striated muscle physiology[J].Arch Biochem Biophys, 2018, 660:121-128.
[30]	GEIST J, KNOTROGIANNI-KONSTANTO-POULOS A.MYBPC1, an emerging myopathic gene:What we know and what we need to learn[J].Front Physiol, 2016, 7:410.
[31]	VELLEMAN S G, COY C S, ABASHT B. Effect of growth selection of broilers on breast muscle satellite cell function:Response of satellite cells to NOV, COMP, MYBP-C1, and CSRP3[J].Comp Biochem Physiol A Mol Integr Physiol, 2021, 255:110917.
[32]	LI D F, PAN Z X, ZHANG K, et al.Identification of the differentially expressed genes of muscle growth and intramuscular fat metabolism in the development stage of yellow broilers[J].Genes (Basel), 2020, 11(3):244.
[33]	XU Y H, DENG J L, WANG L P, et al. Identification of candidate genes associated with breast cancer prognosis[J].DNA Cell Biol, 2020, (7):1205-1227.
[34]	PUDOVA E A, LUKYANOVE E N, NYUSHKO K M, et al.Differentially expressed genes associated with prognosis in locally advanced lymph node-negative prostate cancer[J]. Front Genet, 2019, 10:730.
[35]	STAVUSIS J, LACE B, SCHAFER J, et al.Novel mutations in MYBPC1 are associated with myogenic tremor and mild myopathy[J].Ann of Neurol, 2019, 86(1):129-142.
[36]	ROMANO G D, IBELLI A M, LORENZTTI W R, et al.Inguinal ring RNA sequencing reveals down-regulation of muscular genes related to scrotal hernia in pigs[J].Genes, 2020, 11(2):117.
[37]	GUO W L, GENG J, ZHAO J G, et al.Gene expression profiling reveals upregulated FUT1 and MYBPC1 in children with pancreaticobiliary maljunction[J].Braz J Med Biol Res, 2019, 52(8):e8522.
[38]	WANG X L, LIU Y M, ZHANG Z D, et al.Utilizing benchmarked dataset and gene regulatory network to investigate hub genes in postmenopausal osteoporosis[J].J Cancer Res Ther, 2020, 16(4):867-873.
[39]	VELLEMAN S G, COY C S, ABASHT B. Effect of growth selection of broilers on breast muscle satellite cell function:Response of satellite cells to NOV, COMP, MYBP-C1, and CSRP3[J].Comp Biochem Physiol A Mol Integr Physiol, 2021, 255:110917.

Effect of Overexpression of MyBPC1 on Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells

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