蒙古马两个类群肌纤维发育表型及基因表达谱比较研究

doi:10.11843/j.issn.0366-6964.2025.02.016

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (2): 643-656.doi: 10.11843/j.issn.0366-6964.2025.02.016

蒙古马两个类群肌纤维发育表型及基因表达谱比较研究

胡瀚文¹(), 图格琴¹, 任秀娟¹, 丁文淇¹, 宫文典¹, 贾紫洁¹, 史琳¹, 马木仁², 宝日格乐³, 芒来¹, 白东义¹^,*()

1. 农业农村部马属动物种业创新重点实验室, 内蒙古自治区马属动物科学研究与技术创新重点实验室, 马属动物研究中心, 内蒙古农业大学, 呼和浩特 010018
2. 锡林浩特市农牧技术推广中心, 锡林浩特 026000
3. 科尔沁左翼后旗畜牧水产技术服务中心, 科尔沁 028100

收稿日期:2024-05-20 出版日期:2025-02-23 发布日期:2025-02-26
通讯作者: 白东义 E-mail:havyn0502@qq.com;baidongyi1983@163.com
作者简介:胡瀚文(2000-)，男，山东枣庄人，硕士生，主要从事动物遗传育种研究，E-mail: havyn0502@qq.com
基金资助:
国家自然科学基金(U23A20224)；高校基本科研业务费项目-种业振兴领军人才(BR22-11-03)；农牧业厅种业专项(RK2300003651)；内蒙古自治区科学技术厅自治区科技重大专项(2021ZD0018;2020ZD0004;ZD20190039)

Comparative Study on Muscle Fiber Development Phenotype and Gene Expression Profile of Two Mongolian Horse Populations

HU Hanwen¹(), BAO Tugeqin¹, REN Xiujuan¹, DING Wenqi¹, GONG Wendian¹, JIA Zijie¹, SHI Lin¹, MA Muren², Baorigele ³, DUGARJAVIIN Manglai¹, BAI Dongyi¹^,*()

1. Key Laboratory of Equus Germplasm Innovation of Ministry of Agriculture and Rural Affairs, Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Equus Research Center, Inner Mongolia Agricultural University, Hohhot 010018, China
2. Xilinhot Agricultural and Animal Husbandry Technology Extension Center, Xilinhot 026000, China
3. Horqin Left Back Banner Livestock and Fishery Technology Service Center, Horqin 028100, China

Received:2024-05-20 Online:2025-02-23 Published:2025-02-26
Contact: BAI Dongyi E-mail:havyn0502@qq.com;baidongyi1983@163.com

摘要/Abstract

摘要：

为探究不同类群蒙古马肌肉生长差异，本研究对生长环境差异最大、距离最远的蒙古马类群(巴尔虎马、乌审马)的肌肉表型与肌肉分子层面的差异进行了比较研究。本研究的试验动物分别是3匹生长在陈巴尔虎旗的巴尔虎马和3匹生长在乌审旗的乌审马，均为在野外自由放养采食的健康种公马。每个类群的平均年龄为5岁，同一类群马匹体况相近。其中巴尔虎马宰前活重为(303.10±14.10) kg、胴体重为(148.29±15.43) kg，乌审马宰前活重为(287.90±37.5) kg、胴体重为(140.83±5.04) kg。将试验动物按类群分为2个组，每组3个重复，对所有试验动物进行屠宰并取其臀中肌，将采集的肌肉样本石蜡包埋后进行HE和免疫组化染色，对肌纤维面积以及慢肌纤维占比进行统计；同时对所采集的肌肉样本进行转录组测序，测序结果使用DESeq2软件进行差异基因的筛选，使用David在线软件对差异表达基因进行GO和KEGG富集分析，最后通过qRT-PCR对测序结果进行验证。本试验通过HE染色和免疫组化染色发现，巴尔虎马臀中肌的肌纤维平均面积为(2 592±180.92)μm²，乌审马臀中肌肌纤维面积为(1 997±73.39) μm², 二者差异显著(P < 0.05)；巴尔虎马臀中肌慢肌纤维占比为(10.34±0.59)%，乌审马臀中肌中慢肌纤维占比为(8.14±0.81)%, 二者差异差异不显著(P>0.05)。本研究在巴尔虎马和乌审马的臀中肌中共鉴定出1 103个差异基因，其中有460个上调基因和643个下调基因在乌审马的臀中肌中表达。研究发现，MYH15、MYOZ2、多个谷氨酸受体基因和多个GABAA型受体基因在巴尔虎马臀中肌中高表达；MYH6和FOXO1基因在乌审马臀中肌中高表达。对差异基因进行富集分析发现，GO分析富集到了208个条目，KEGG分析富集到了65个通路，所富集到的条目与通路主要和谷氨酸信号传导、GABA信号传导以及肌肉生长发育有关。本试验对巴尔虎马和乌审马的臀中肌进行研究，发现在二者臀中肌肌肉表型方面，巴尔虎马肌纤维面积显著大于乌审马，但慢肌纤维占比差异不显著；在分子层面二者有较为明显的差异，这些差异集中在了肌纤维类型、肌纤维面积以及肌肉中肌梭神经信号传导等方面。

关键词: 巴尔虎马, 乌审马, 肌纤维面积, 转录组

Abstract:

To investigate the differences in muscle growth among different populations of Mongolian horses, the muscle phenotype and molecular-level differences between the Mongolian horse populations with the most significant environmental differences and the greatest geographical distance(Baerhu horse and Wushen horse) was compared. The experimental animals in this study consisted of 3 Baerhu horses raised in the Chenbaerhu Banner and 3 Wushen horses raised in the Wushen Banner. All horses were healthy stallions, free-range grazing in the wild. The average age of the horses in each population was 5 years, with similar body conditions within each population. The pre-slaughter live weight of the Baerhu horses was (303.10±14.10) kg, and the carcass weight was (148.29±15.43) kg. The pre-slaughter live weight of the Wushen horses was (287.90±37.5) kg, and the carcass weight was (140.83±5.04) kg. The experimental animals were divided into two populations based on their breed, with 3 repetitions per group. All animals were slaughtered, and their gluteus medius muscles were collected. After the muscle samples were embedded in paraffin, HE and immunohistochemistry staining were performed. The muscle fiber area and the proportion of slow-twitch muscle fibers were statistically analyzed. Additionally, transcriptome sequencing was conducted on the collected muscle samples. The sequencing data were analyzed for differentially expressed genes using the DESeq2 software, and GO and KEGG enrichment analyses were performed using the David online tool. Finally, qRT-PCR was used to validate the sequencing results. In this experiment, HE staining and immunohistochemical staining revealed that the average muscle fiber area of the gluteus medius in Baerhu horses was (2 592±180.92) μm², while in Wushen horses it was (1 997±73.39) μm², with a significant difference between the two populations (P < 0.05). The proportion of slow-twitch muscle fibers in the gluteus medius of Baerhu horses was (10.34±0.59)%, while in Wushen horses it was (8.14±0.81)%, with no significant difference between the two populations (P>0.05).This study identified a total of 1 103 differentially expressed genes in the gluteus medius of Baerhu and Wushen horses, with 460 upregulated genes and 643 downregulated genes expressed in the gluteus medius of Wushen horses. The study found that MYH15, MYOZ2, several glutamate receptor genes, and several GABAA receptor genes were highly expressed in the gluteus medius of Baerhu horses, while MYH6 and FOXO1 genes were highly expressed in the gluteus medius of Wushen horses. Enrichment analysis of the differentially expressed genes revealed 208 enriched items in the GO analysis and 65 pathways in the KEGG analysis. These enriched items and pathways were mainly associated with glutamate signaling, GABA signaling, and muscle growth and development. This study investigates the gluteus medius muscle of the Baerhu horse and the Wushen horse, finding that the fiber area of the gluteus medius in Baerhu horses is significantly larger than that in Wushen horses, while the proportion of slow muscle fibers shows no significant difference. At the molecular level, there are notable differences between the two populations, particularly in muscle fiber types, fiber area, and neuromuscular signaling in the muscle spindles.

Key words: Baerhu horse, Wushen horse, muscle fiber area, transcriptome

中图分类号:

S821.2

胡瀚文, 图格琴, 任秀娟, 丁文淇, 宫文典, 贾紫洁, 史琳, 马木仁, 宝日格乐, 芒来, 白东义. 蒙古马两个类群肌纤维发育表型及基因表达谱比较研究[J]. 畜牧兽医学报, 2025, 56(2): 643-656.

HU Hanwen, BAO Tugeqin, REN Xiujuan, DING Wenqi, GONG Wendian, JIA Zijie, SHI Lin, MA Muren, Baorigele , DUGARJAVIIN Manglai, BAI Dongyi. Comparative Study on Muscle Fiber Development Phenotype and Gene Expression Profile of Two Mongolian Horse Populations[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(2): 643-656.

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基因 Gene	引物序列(5'→3') Primer sequence
IVD	F：GGAAGATGGCTGACATGTACA
	R：GCTGAGTAAAGAATCACCCCT
PEAR1	F：TTCACCATAATGCCTACCTCTC
	R：GGATATCTGGCATGACGTACTC
MYH6	F：TACCAGTACATGCTGACAGATC
	R：CAAAGTACTGGATGACACGTTT
NYAP2	F：CTACCCTAAAAGTCACTCGGC
	R：GGTAAAGAGGCTGGTTAGTTCT
SYT4	F：GAGTTCTGAGAAAGACCTTGGA
	R：AAAACTCAAGATCGTGAAGTGC
CALML5	F：CCTTCTCCAGTGTTGACAAGAA
	R：GAACTCTTGGAAGTTGATGACG
GAPDH	F：CATCATCCCTGCTTCTACTGG
	R：TCCACGACTGACACGTTAGG

蒙古马类群 Mongolian horse populations	平均肌纤维面积±标准误/μm² Average muscle fiber area±standard error	平均慢肌纤维占比±标准误/% Average proportion of slow muscle fibers±standard error
巴尔虎马Baerhu horse	2 592±180.92^a	10.34±0.59^a
乌审马Wushen horse	1 997±73.39^b	8.14±0.81^a

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蒙古马两个类群肌纤维发育表型及基因表达谱比较研究

Comparative Study on Muscle Fiber Development Phenotype and Gene Expression Profile of Two Mongolian Horse Populations

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