胎儿期与成年期蒙古马骨骼肌肌纤维类型转化研究

doi:10.11843/j.issn.0366-6964.2022.01.009

畜牧兽医学报 ›› 2022, Vol. 53 ›› Issue (1): 88-99.doi: 10.11843/j.issn.0366-6964.2022.01.009

胎儿期与成年期蒙古马骨骼肌肌纤维类型转化研究

丁文淇¹, 图格琴¹, 任秀娟¹, 陶克涛¹, 拉希玛¹, 贾紫洁¹, 安塔娜¹, 铁木齐尔·阿尔滕齐米克¹, 韩海格¹, 陶娜拉², 芒来^1*, 白东义^1*

1. 内蒙古农业大学动物科学学院, 内蒙古自治区马遗传育种与繁殖重点实验室, 农业农村部马属动物遗传育种繁殖科学观测实验站, 内蒙古农业大学马属动物研究中心, 呼和浩特 010018;
2. 锡林郭勒盟镶黄旗农牧和科技局, 镶黄旗 013250

收稿日期:2021-04-29 出版日期:2022-01-23 发布日期:2022-01-26
通讯作者: 芒来,主要从事马属动物遗传育种与繁殖研究,E-mail:dmanglai@163.com;白东义,主要从事马属动物遗传育种与繁殖研究,E-mail:baidongyi1983@163.com
作者简介:丁文淇(1997-),男,内蒙古赤峰人,硕士生,主要从事动物遗传育种研究,E-mail:2510060080@qq.com
基金资助:
国家自然科学基金（31960657；31961143025）；内蒙古自治区科技重大专项（ZD20190039）；内蒙古自治区自然科学基金项目（2019MS03064）；内蒙古农业大学动物科学学院标志性成果专项资金项目（BZCG202115）

Study on Muscle Fiber Type Transformation of Mongolian Horse during Fetal and Adult Period

DING Wenqi¹, BAO Tugeqin¹, REN Xiujuan¹, MONGKE Togtokh¹, LA Xima¹, JIA Zijie¹, AN Tana¹, TIEMUQIER Aertengqimike¹, HAN Haige¹, TAO Nala², DUGARJAVIIN Manglai^1*, BAI Dongyi^1*

1. Equine Research Center, Scientific Observing and Experimental Station of Equine Genetics, Breeding and Reproduction of Ministry of Agriculture and Rural Affairs, Inner Mongolia Key Laboratory of Equine Genetics, Breeding and Reproduction, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China;
2. Xilingol Leagne Xianghuangqi Agriculture, Animal Husbandry and Science and Technology Bureau, Xianghuangqi 013250, China

Received:2021-04-29 Online:2022-01-23 Published:2022-01-26

摘要/Abstract

摘要： 为探索影响蒙古马胎儿期和成年期肌纤维类型差异机理。本研究选取3匹4月龄胎儿（两母一公）与3匹5岁健康成年母马身体4块分布全身、具有代表性的肌肉组织（长臂三头肌、夹肌、背最长肌、臀中肌）作为一个整体。胎儿期蒙古马肌纤维和成年期蒙古马肌纤维因存在差异各做为一组，试验进行3个生物学重复。首先对蒙古马骨骼肌肌肉样品进行免疫组化染色观察，接下来对骨骼肌样品进行RNA-seq，构建胎儿期和成年期蒙古马不同时期肌纤维mRNA的表达谱。筛选差异表达基因，进行GO和KEGG功能富集分析。组织学结果显示，胎儿期快肌纤维占比显著高于成年期，RNA-seq筛选可能影响肌纤维类型转化的候选基因，比较胎儿期和成年期蒙古马影响肌纤维类型的基因表达谱。共筛出250个差异表达基因，在成年期上调的基因有27个，包括TNNT1、MYOZ2、ATP2A2等，在胎儿期上调的基因17个，包括MYL1、TNNT3、ENO3等。其中ATP2A2和TNNT1主要与快、慢肌纤维类型转化有关，这些差异表达基因还与骨骼肌生长、发育密切相关。结果表明，胎儿期蒙古马快肌纤维比例显著高于成年期蒙古马。ATP2A2、MYOZ2等基因主要表达在成年期蒙古马肌纤维，ENO3、TNNI2等基因主要表达在胎儿期蒙古马肌纤维。钙信号和AMPK信号通路与肌纤维类型转化有关。本试验结果对于研究蒙古马肌纤维类型转化具有重要意义。

关键词: 蒙古马, 转录组, 肌纤维类型

Abstract: The study aimed to explore the mechanism of muscle fiber type difference between fetal and adult Mongolian horses. In this study, three 4-month-old fetuses (two females and one male) and three 5-year-old healthy adult female horses were selected as a whole with 4 body-distributed and representative muscle tissues (triceps brachii, splenius, longissimus dorsi and gluteus medius). Fetal Mongolian horse muscle fiber was one group, and adult Mongolian horse muscle fiber was one group, the experiment carried out 3 biological repetitions.Firstly, the skeletal muscle samples of Mongolian horses were observed by immunohistochemical staining. Then, the skeletal muscle samples were subjected to RNA-seq to construct the mRNA expression profiles of skeletal muscle fibers in different periods of fetal and adult Mongolian horses. Differentially expressed genes were screened for GO and KEGG functional enrichment analysis. Histological results showed that the proportion of fast muscle fibers in the fetal period was significantly higher than that in the adult period. RNA-seq was used to screen candidate genes that might affect the transformation related to muscle fiber types. The gene expression profiles related to muscle fiber types in fetal and adult Mongolian horses were compared. A total of 250 differentially expressed genes were screened out. There were 27 up-regulated genes in adulthood, including TNNT1, MYOZ2 and ATP2A2, and 17 up-regulated genes in the fetal period, including MYL1, TNNT3 and ENO3. ATP2A2 and TNNT1 were mainly related to fast and slow muscle fiber type transformation, and these differentially expressed genes were also closely related to skeletal muscle growth and development. The results showed that the proportion of fast muscle fibers in fetal Mongolian horses was significantly higher than that in adult Mongolian horses. ATP2A2, MYOZ2 were mainly expressed in muscle fibers of adult Mongolian horse, ENO3, TNNI2 were mainly expressed in muscle fibers of fetal Mongolian horse. Calcium and AMPK signaling pathways are related to muscle fiber type conversion.The results of this experiment are of great significance to the study of Mongolian horse muscle fiber type transformation.

Key words: Mongolian horse, transcriptome, muscle fiber type

中图分类号:

S821.2

丁文淇, 图格琴, 任秀娟, 陶克涛, 拉希玛, 贾紫洁, 安塔娜, 铁木齐尔·阿尔滕齐米克, 韩海格, 陶娜拉, 芒来, 白东义. 胎儿期与成年期蒙古马骨骼肌肌纤维类型转化研究[J]. 畜牧兽医学报, 2022, 53(1): 88-99.

DING Wenqi, BAO Tugeqin, REN Xiujuan, MONGKE Togtokh, LA Xima, JIA Zijie, AN Tana, TIEMUQIER Aertengqimike, HAN Haige, TAO Nala, DUGARJAVIIN Manglai, BAI Dongyi. Study on Muscle Fiber Type Transformation of Mongolian Horse during Fetal and Adult Period[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(1): 88-99.

参考文献 38

[1]	赵启南.蒙古马高负荷运动训练前后转录组与miRNA文库特征及关联分析[D].呼和浩特:内蒙古农业大学,2015.ZHAO Q N.Mongolian horse transcriptome and miRNA library characteristics and correlation analysis before and after high load exercise training[D].Hohhot:Inner Mongolia Agricultural University,2015.(in Chinese)
[2]	欧秀琼,李睿,张晓春,等.猪肌纤维性状形成和肌内脂肪沉积的遗传机制[J].中国畜牧兽医,2021,48(3):925-931.OU X Q,LI R,ZHANG X C,et al.Genetic mechanism of pig muscle fiber property formation and intramuscular fat deposition[J]. China Animal Husbandry & Veterinary Medicine,2021,48(3):925-931.(in Chinese)
[3]	SCHIAFFINO S,REGGIANI C.Fiber types in mammalian skeletal muscles[J].Physiol Rev,2011,91(4):1447-1531.
[4]	任阳.饱和与不饱和脂肪酸对猪肌纤维组成的影响及其AMPK途径研究[D].杭州:浙江大学,2014:7-8.REN Y.Effect of saturated and unsaturated fatty acids on porcine muscle fiber composition and AMPK expression[D]. Hangzhou:Zhejiang University,2014:7-8.(in Chinese)
[5]	孙一,朱荣,梁永桥,等.高强度间歇训练对不同类型肌纤维代谢与分布的调节[J].广州体育学院学报,2018,38(1):82-89.SUN Y,ZHU R,LIANG Y Q,et al.Regulation of high intensity interval training on metabolism and distribution of different fiber-type skeletal muscle[J].Journal of Guangzhou Sport University,2018,38(1):82-89.(in Chinese)
[6]	郭秋平,文超越,王文龙,等.肌纤维类型转化的分子信号通路及其营养调控进展[J].动物营养学报,2017,29(6):1836-1842.GUO Q P,WEN C Y,WANG W L,et al.Progress in molecular signaling pathway of skeletal muscle fiber type transformation and its nutritional regulation[J].Chinese Journal of Animal Nutrition,2017,29(6):1836-1842.(in Chinese)
[7]	PICARD B,BERRI C,LEFAUCHEUR L,et al.Skeletal muscle proteomics in livestock production[J].Brief Funct Genomics,2010, 9(3): 259-278.
[8]	SAKUMA K,WATANABE K,SANO M,et al.The adaptive response of MyOD family proteins in overloaded,regenerating and denervated rat muscles[J].Biochim Biophys Acta-Gen Subj,1999,1428(2-3):284-292.
[9]	ALLEN D L,SARTORIUS C A,SYCURO L K,et al.Different pathways regulate expression of the skeletal myosin heavy chain genes.[J].J Biol Chem,2001,276(47):43524-43533.
[10]	MURRE C,MCCAW P S,VAESSIN H,et al.Interactions between heterologous helix-loop-helix proteins generate complexes that bind specifically to a common DNA sequence[J].Cell,1989,58(3):537-544.
[11]	牛姣艳.Pax7、MyOD和MyoG基因在猪背最长肌中的发育性表达研究[D].晋中:山西农业大学,2015.NIU J Y.Study on the developmental expression of Pax7,MyoD and MyoG genes in longissimus dorsi of pigs[D].Jinzhong:Shanxi Agricultural University,2015.(in Chinese)
[12]	EKMARK M,RANA Z A,STEWART G,et al.De-phosphorylation of MyOD is linking nerve-evoked activity to fast myosin heavy chain expression in rodent adult skeletal muscle[J].J Physiol,2007,584(2):637-650.
[13]	MACHARIA R,OTTO A,VALASEK P,et al.Neuromuscular junction morphology,fiber-type proportions,and satellite-cell proliferation rates are altered in MyOD-/- mice[J].Muscle Nerve,2010,42(1):38-52.
[14]	ALAPAT D V,CHAUDHRY T,ARDAKANY-TAGHAVI R,et al.Fiber-types of sarcomeric proteins expressed in cultured myogenic cells are modulated by the dose of myogenin activity[J].Cell Signal,2009,21(1):128-135.
[15]	王琼,朱庆.肌肉生长相关因子MyoG基因的研究进展[J].黑龙江畜牧兽医,2007(4):23-24.WANG Q,ZHU Q.Research progress of MyoG gene related to muscle growth[J].Heilongjiang Animal Science and Veterinary Medicine,2007(4):23-24.(in Chinese)
[16]	MORESI V,WILLIAMS A H,MEADOWS E,et al.Myogenin and class II HDACs control neurogenic muscle atrophy by inducing E3 ubiquitin ligases[J].Cell,2010,143(1):35-45.
[17]	PERTEA M,KIM D,PERTEA G M,et al.Transcript-level expression analysis of RNA-seq experiments with HISAT,StringTie and Ballgown[J].Nat Protoc,2016,11(9):1650-1667.
[18]	LOVE M I,HUBER W,ANDERS S.Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2[J].Genome Biol,2014,15(12):550.
[19]	白艳苹,侯艳茹,苏琳,等.乳酸菌诱导线粒体生物发生对绵羊肌纤维特性和肉品质的影响[J].农业工程学报,2021,37(10): 269-276.BAI Y P,HOU Y R,SU L,et al.Effects of lactobacillus induced mitochondrial biogenesis on muscle fiber properties and meat quality of sheep[J].Transactions of the Chinese Society of Agricultural Engineering,2021,37(10):269-276.(in Chinese)
[20]	朱文奇,徐文娟,束婧婷,等.鸭骨骼肌早期发育过程中钙蛋白酶3(CAPN3)基因的表达及其与肌纤维性状的关联[J].畜牧兽医学报, 2014,45(3):385-390.ZHU W Q,XU W J,SHU J T,et al.Expression of CAPN3 gene in skeletal muscles and its association with myofiber traits during embryonic and early post-hatching development in ducks[J].Acta Veterinaria et Zootechnica Sinica,2014,45(3):385-390.(in Chinese)
[21]	EIZEMA K,VAN DER WAL D E,VAN DEN BURG M M M,et al.Differential expression of calcineurin and SR Ca2+ handling proteins in equine muscle fibers during early postnatal growth[J].J Histochem Cytochem,2007,55(3):247-254.
[22]	李玥,袁丽霞,杨晓静,等.早期和后期限饲对肉鸡腓肠肌肌纤维类型和生长轴相关基因表达的影响[J].动物学报,2006,52(6):1133-1141.LI Y,YUAN L X,YANG X J,et al.Effects of early and late feed restriction on myofiber types and expression of growth-related genes in gastrocnemius of broiler chickens[J].Acta Zoologica Sinica,2006,52(6):1133-1141.(in Chinese)
[23]	BAO T,HAN H G,LI B,et al.The distinct transcriptomes of fast-twitch and slow-twitch muscles in Mongolian horses[J].Comp Biochem Physiol Part D:Genomics Proteomics,2020,33:100649.
[24]	PETTE D,STARON R S.Myosin isoforms,muscle fiber types,and transitions[J].Microsc Res Tech,2000,50(6):500-509.
[25]	SWOAP S J,HUNTER R B,STEVENSON E J,et al.The calcineurin-NFAT pathway and muscle fiber-type gene expression[J].Am J Physiol Cell Physiol,2000,279(4):C915-C924.
[26]	字向东,罗斌,夏威,等.基于RNA-Seq技术的牦牛体外受精胚胎发育转录组分析[J].中国农业科学,2018,51(8): 1577-1589.ZI X D,LUO B,XIA W,et al.Transcriptomic analysis of IVF embryonic development in the yak (Bos grunniens) via RNA-Seq[J].Scientia Agricultura Sinica,2018,51(8):1577-1589.(in Chinese)
[27]	SCHIAFFINO S,SANDRI M,MURGIA M.Activity-dependent signaling pathways controlling muscle diversity and plasticity[J]. Physiology (Bethesda),2007,22(4):269-278.
[28]	THOMSON D M.The role of AMPK in the regulation of skeletal muscle size,hypertrophy,and regeneration[J].Int J Mol Sci,2018,19(10):3125.
[29]	王慧婷,张岩晨,徐梦怡,等.能量代谢关键调控因子PGC-1α的研究进展[J].生理学报,2020,72(6):804-816.WANG H T,ZHANG Y C,XU M Y,et al.Research progresses on PGC-1α,a key energy metabolic regulator[J].Acta Physiologica Sinica,2020,72(6):804-816.(in Chinese)
[30]	王亚宁.MEF2A对秦川牛骨骼肌成肌细胞增殖和分化的调控作用及机理研究[D].咸阳:西北农林科技大学,2019.WANG Y N.The Roles of MEF2A in the regulation of skeletal muscle myoblasts proliferation and differentiation in Qinchuan Beef cattle[D].Xianyang:Northwest A&F University,2019.(in Chinese)
[31]	FREY N,RICHARDSON J A,OLSON E N.Calsarcins,a novel family of sarcomeric calcineurin-binding proteins[J].Proc Natl Acad Sci U S A,2000,97(26):14632-14637.
[32]	FREY N,BARRIENTOS T,SHELTON J M,et al.Mice lacking calsarcin-1 are sensitized to calcineurin signaling and show accelerated cardiomyopathy in response to pathological biomechanical stress[J].Nat Med,2004,10(12):1336-1343.
[33]	RUGGIERO A,CHEN S N,LOMBARDI R,et al.Pathogenesis of hypertrophic cardiomyopathy caused by myozenin 2 mutations is independent of calcineurin activity[J].Cardiovasc Res,2013,97(1):44-54.
[34]	王恒.肌肉中Calsarcin基因的差异表达和转录调控研究[D].武汉:华中农业大学,2007.WANG H.Characterization of the different expression patterns and transcription regulations of Calsarcins in muscle[D]. Wuhan:Huazhong Agricultural University,2007.(in Chinese)
[35]	ELDOR R,NORTON L,FOURCAUDOT M,et al.Increased lipid availability for three days reduces whole body glucose uptake,impairs muscle mitochondrial function and initiates opposing effects on PGC-1α promoter methylation in healthy subjects[J].PLoS One,2017,12(12):e0188208.
[36]	张鑫,王宇波,黄志清,等.猪肉品质营养调控的研究进展[J].动物营养学报,2020,32(10):4555-4564.ZHANG X,WANG Y B,HUANG Z Q,et al.Research progress of nutritional regulation for meat quality of pigs[J].Chinese Journal of Animal Nutrition,2020,32(10):4555-4564.(in Chinese)
[37]	曾子悠.饲粮添加白藜芦醇对断奶仔猪肌纤维类型、抗氧化能力及肠道屏障功能的影响[D].雅安:四川农业大学,2019.ZENG Z Y.Effect of dietary resveratrol supplementation on skeletal muscle fiber type,antioxidant capacity and intestinal barrier function in weaned piglets[D].Ya’an:Sichuan Agricultural University,2019.(in Chinese)
[38]	欧秀琼,李星,钟正泽,等.猪肌肉肌纤维特性与肌肉品质的关系及品种、性别差异[J].新疆农业科学,2019,56(12):2345-2352.OU X Q,LI X,ZHONG Z Z,et al.Relationship of muscle fiber characteristics with meat quality and its breed and sex differences in pigs[J].Xinjiang Agricultural Sciences,2019,56(12):2345-2352.(in Chinese)

胎儿期与成年期蒙古马骨骼肌肌纤维类型转化研究

Study on Muscle Fiber Type Transformation of Mongolian Horse during Fetal and Adult Period

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献 38

相关文章 15

编辑推荐

Metrics

本文评价

[1]	刘益丽, 唐娇, 闵奇, 杨露, 王泽宁, 胡莲, 赵迪, 江明锋. 基于转录组数据挖掘牦牛皱胃发育代谢的关键候选基因[J]. 畜牧兽医学报, 2024, 55(1): 153-168.
[2]	梁凯欣, 钟海文, 宋长绪, 杨化强, 黄思秀, 徐铮. SYNGR2影响猪圆环病毒2型体外增殖的研究[J]. 畜牧兽医学报, 2023, 54(9): 3824-3835.
[3]	胡婷, 张永红, 侯晓林, 姚华, 崔德凤, 潘早早, 张凌宇, 张家希, 吴琼. 基于转录组学研究双酚A对猪睾丸支持细胞炎症和氨基酸代谢通路的影响[J]. 畜牧兽医学报, 2023, 54(7): 2858-2871.
[4]	刘航, 王欢欢, 葛莹, 张雷, 张伟武, 魏莹晖, 李庆海, 范京辉, 章学东. 基于转录组和蛋白组筛选乌骨鸡肤色性状候选基因[J]. 畜牧兽医学报, 2023, 54(6): 2320-2329.
[5]	白露, 王梦杰, 马小春, 何政肖, 孔富丽, 刘大伟, 营凡, 朱丹, 赵桂苹, 文杰, 刘冉冉. 鸡木质化胸肌组织学特征及分子调控通路改变研究[J]. 畜牧兽医学报, 2023, 54(5): 1915-1926.
[6]	王美慧, 钟震宇, 白加德, 单云芳, 程志斌, 张庆勋, 孟玉萍, 董玉兰, 郭青云. C型产气荚膜梭菌感染鹿肠道中关键基因和途径的转录组分析[J]. 畜牧兽医学报, 2023, 54(5): 2147-2157.
[7]	孙美杰, 曹力文, 郑文金, 申军士, 朱伟云. 基于转录组学研究日粮添加尿素对育肥湖羊肝组织氨代谢的影响[J]. 畜牧兽医学报, 2023, 54(3): 1148-1159.
[8]	刘源壹, 李昕俞, 巴音那木拉, 翠芳, 芒来, 杜明. 单细胞转录组测序技术及其在动物繁殖中的应用进展[J]. 畜牧兽医学报, 2023, 54(2): 421-433.
[9]	张高猛, 丁纪强, 刘昱宏, 郑麦青, 文杰, 赵桂苹, 李庆贺. 全基因组关联分析揭示白羽肉鸡孵化性状的遗传基础[J]. 畜牧兽医学报, 2023, 54(2): 534-544.
[10]	贾紫洁, 图格琴, 丁文淇, 任秀娟, 刘慧莹, 李欣泽, 翠芳, 芒来, 白东义. 蒙古马全身主要骨骼肌表型谱的构建及比较研究[J]. 畜牧兽医学报, 2023, 54(2): 596-607.
[11]	余祖华, 贾艳艳, 何雷, 廖成水, 李静, 魏颖, 陈建, 陈松彪, 尚珂, 丁轲. gga-miR-155对MDCC-MSB1细胞转录组的影响[J]. 畜牧兽医学报, 2023, 54(2): 663-672.
[12]	任秀娟, 康德措, 苏少锋, 蔺雅楠, 李雅静, 杜明, 白东义, 李蓓, 赵一萍, 芒来. 蒙古马和纯血马免疫相关基因差异表达分析[J]. 畜牧兽医学报, 2023, 54(12): 5020-5032.
[13]	韩浩园, 李世凯, 杨瑞巧, 李曼曼, 李君, 哈斯, 赵金艳, 魏红芳, 权凯. 基于转录组测序挖掘槐山羊高繁关键候选基因[J]. 畜牧兽医学报, 2023, 54(12): 5077-5090.
[14]	王文翔, 杜丽丽, 胡俊伟, 张琰翔, 马铭昊, 段蕊, 钱聪, 王欣玥, 李三禄, 张长庆, 张路培, 高雪, 徐凌洋, 李俊雅, 高会江. 基于转录组分析挖掘平凉红牛背最长肌肉质性状候选基因[J]. 畜牧兽医学报, 2023, 54(11): 4589-4604.
[15]	张倩, 崔燕, 余四九, 何俊峰, 潘阳阳, 王萌. 初生及成年牦牛大脑皮质的转录组分析[J]. 畜牧兽医学报, 2023, 54(11): 4605-4614.