中国畜禽种业 ›› 2026, Vol. 22 ›› Issue (5): 32-42.doi: 10.19543/j.cnki.1673-4556.20260416.001

• 生物技术 • 上一篇    下一篇

信阳水牛肌纤维类型调控相关基因组织表达及分化时序研究

张潇戈1(), 周艳朵2, 李建彰1, 马路平1, 李军1, 郝瑞杰3, 张子敬4, 马云5, 张天留6, 梁成成1()   

  1. 1.信阳农林学院动物科技学院，河南 信阳 464000
    2.信阳农林学院农学院，河南 信阳 464000
    3.信阳师范大学生命科学学院，河南 信阳 464000
    4.河南省农业科学院畜牧兽医研究所，河南 郑州 450000
    5.宁夏大学动物科技学院，宁夏 银川 750000
    6.河南农业大学动物科技学院，河南 郑州 450000
  • 收稿日期:2025-11-26 出版日期:2026-05-26 发布日期:2026-06-17
  • 通讯作者: 梁成成 E-mail:OnlyIvan2435@163.com;lcc20151120@xyafu.edu.cn
  • 作者简介:张潇戈（2003—），女，河南郑州人，研究方向：反刍动物健康养殖，E-mail：OnlyIvan2435@163.com。
    张潇戈，信阳农林学院动物科技学院，本科生，参与项目有河南省自然科学基金青年基金项目（252300423641）、河南省大学生创新创业训练计划（202511326001）以及信阳农林学院本科生科研训练项目（URPDK202501），曾获校三好学生，校优秀团员，校极优秀学生组织干部等荣誉。
    梁成成，农学博士，信阳农林学院校聘副教授，主要研究方向水牛分子育种与健康养殖；兼任中国农学会秸秆资源综合利用分会理事、河南省“科技副总”、《中国畜禽种业》青年编委，获校级及以上荣誉9项；主持校级以上项目2项，参与省部级及以上课题4项；近5年累计发表各类论文31篇，获国家专利1项，参与制定地方标准1项，在信阳水牛研究领域形成扎实科研基础与系列成果。
  • 基金资助:
    河南省自然科学基金青年基金(252300423641);河南省大学生创新创业训练计划(202511326001);信阳农林学院本科生科研训练项目(URPDK202501)

Tissue expression and differentiation characteristics of genes regulating muscle fiber types in Xinyang buffalo

Xiaoge Zhang1(), Yanduo Zhou2, Jianzhang Li1, Luping Ma1, Jun Li1, Ruijie Hao3, Zijing Zhang4, Yun Ma5, Tianliu Zhang6, Chengcheng Liang1()   

  1. 1.College of Animal Science and Technology, Xinyang Agricultural and Forestry University, Xinyang, 464000, Henan
    2.College of Agronomy, Xinyang Agricultural and Forestry University, Xinyang, 464000, Henan
    3.College of Life Sciences, Xinyang Normal University, Xinyang, 464000, Henan
    4.Institute of Animal Husbandry and Veterinary Medicine, Henan Academy of Agricultural Sciences, Zhengzhou, 450000, Henan
    5.College of Animal Science and Technology, Ningxia University, Yinchuan, 750000, Ningxia
    6.College of Animal Science and Technology, Henan Agricultural University, Zhengzhou, 450000, Henan
  • Received:2025-11-26 Online:2026-05-26 Published:2026-06-17
  • Contact: Chengcheng Liang E-mail:OnlyIvan2435@163.com;lcc20151120@xyafu.edu.cn

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摘要：

目的 本研究旨在解析信阳水牛中与慢肌纤维向快肌纤维转化相关的肌肉因子MSTN、BDNF和DKK3的功能及其对肉质特性的影响，以期为信阳水牛品种改良提供参考。 方法 根据相关文献选取MSTN、BDNF、DKK3三个肌纤维因子，利用NCBI下载其氨基酸序列，利用MEME进行结构域预测，利用在线软件STRING分析蛋白互作情况，对其进行生物信息学分析。采集3头36月龄公牛的肝脏、脾脏、肺脏、肾脏、心脏、前腿肌、后腿肌、背最长肌、皮下肌、大肠、小肠等11种组织同时进行组织表达谱分析。采集1头1日龄新生牛的背最长肌，进行分离培养，诱导其分化并进行时序表达。 结果 生物学信息学分析显示，MSTN基因编码的蛋白质含有TGF-β信号通路相关结构域，主要在骨骼肌组织中高表达。BDNF基因编码的蛋白质含有神经营养因子相关结构域，主要在神经相关组织和骨骼肌中高表达。DKK3基因编码的蛋白质含有Wnt信号通路相关结构域，主要在脂肪和肝脏组织中高表达。GO和KEGG分析进一步表明，这些基因分别富集于TGF-β信号通路、神经营养因子信号通路和Wnt信号通路。时序分化结果表明，MSTN在分化前期表达上调，随后快速下调；BDNF基因表达量呈逐渐上升的趋势，在分化晚期再次被激活上调；DKK3在原代成肌细胞分化过程中不表达。 结论 以上结果表明MSTN、BDNF、DKK3基因表达与水牛肌纤维的形成存在关联。

关键词: 信阳水牛, MSTN, BDNF, DKK3, 生物信息学分析, 组织表达

Abstract:

Objective To elucidate the functions of MSTN, BDNF, and DKK3 genes associated with slow-to-fast muscle fiber conversion in Xinyang buffalo and their effects on meat quality characteristics, in order to provide reference for the improvement of Xinyang buffalo breeds. Method Three muscle fiber-related genes (MSTN, BDNF, DKK3) were selected from relevant literature. Their amino acid sequences were downloaded from NCBI, domain prediction was performed using MEME, and protein interaction analysis was conducted through the online software STRING for bioinformatics evaluation. Tissue samples from three 36-month-old bulls (Liver, spleen, lung, kidney, heart, foreleg muscle, hindleg muscle, longissimus dorsi muscle, subcutaneous muscle, large intestine, small intestine) were collected for tissue expression profiling. Collect the longissimus dorsi muscle from a 1-day-old newborn calf, isolate and culture the primary cells, induce their differentiation, and analyze time-course gene expression. Result Bioinformatics analysis reveals that the protein encoded by the MSTN gene contains domains associated with the TGF-β signaling pathway and is predominantly highly expressed in skeletal muscle tissue. The protein encoded by the BDNF gene possesses domains related to neurotrophic factors and is primarily highly expressed in neural tissues and skeletal muscle. The protein encoded by the DKK3 gene contains domains associated with the Wnt signaling pathway and is mainly highly expressed in adipose and liver tissues. GO and KEGG analyses further indicated that these genes were respectively enriched in the TGF-β signaling pathway, neurotrophic factor signaling pathway, and Wnt signaling pathway. The temporal differentiation results showed that the expression of MSTN was upregulated in the early stage of differentiation and then rapidly downregulated; the expression level of the BDNF gene exhibited a gradual upward trend and was reactivated and upregulated at the late differentiation stage; DKK3 was not expressed during the differentiation of primary myoblasts. Conclusion These results indicate that the expression of MSTN, BDNF, and DKK3 genes is associated with the formation of muscle fibers in buffalo.

Key words: Xinyang buffalo, MSTN, BDNF, DKK3, Bioinformatics analysis, Tissue expression

中图分类号: 

  • S823

表1

MSTN、BDNF及DKK3基因的引物信息"

基因Genes引物序列（5'-3'）Primer sequence产物长度Product length/bp退火温度Annealing temperature/℃
MSTN-FCATTACCATGCCCACGGAGTCTG14963.6
MSTN-RTCGCAGGAGTCTTGACAGGTCTC
BDNF-FAACTCCCAGTGCCGAACTACCC8064.1
BDNF-RATGAACCGCCAGCCAATACGC
DKK3-FTCGTCCTCTCGGCAGTGTTACTC14863.6
DKK3-RACTCGGGCAAGCAAGATGACATC
β-actin-FCATCCTGACCCTCAAGTA9152.3
β-actin-RCTCGTTGTAGAAGGTGTG

图1

基于MSTN、BDNF及DKK3基因的氨基酸序列motif分析"

图2

基于MSTN、BDNF及DKK3基因的氨基酸序列结构域分析"

图3

MSTN、BDNF、DKK3蛋白互作预测结构图"

表2

蛋白互作网络KEGG富集分析"

核心蛋白

Core protein

通路ID

Term ID

通路描述

Term description

基因数量

Gene count

错误率

FDR

基因

Genes

MSTNbta04350TGF-β信号通路67.56E-10FST, SMAD2, TGFBR1, ACVR2A, ACVR1B, ACVR2B
bta04550调节干细胞多能性56.67E-07SMAD2, ACVR2A, MYF5, ACVR1B, ACVR2B
bta04060细胞因子-受体相互作用51.62E-05MSTN, TGFBR1, ACVR2A, ACVR1B, ACVR2B
BDNFbta04722神经营养因子62.24E-09BDNF, NGFR, NTRK2, NTF4, NGF, SORT1
bta04151PI3K-Akt信号通路69.36E-07BDNF, IL6, NGFR, NTRK2, NTF4, NGF
bta04014Ras信号通路55.85E-06BDNF, NGFR, NTRK2, NTF4, NGF
bta04010MAPK信号通路51.03E-05BDNF, NGFR, NTRK2, NTF4, NGF
DKK3bta04310Wnt信号通路76.33E-11LRP5, DKK2, SFRP4, CTNNB1, SFRP2, SFRP1, DKK1

表3

蛋白互作网络GO富集分析"

项目

Items

基因

Genes

项目ID

Term ID

项目描述

Term description

基因数量

Gene count

错误率FDR

基因

Genes

细胞组分

Cellular component

MSTNGO: 0048179激活素受体复合体41.99E-08TGFBR1, ACVR2A, ACVR1B, ACVR2B
BDNFGO: 0045202突触50.0174BDNF, NGFR, NTRK2, NTF4, NGF
GO: 0120025质膜界定的细胞突起60.0174CNTF, BDNF, NTRK2, NTF4, NGF, GFAP
DKK3GO: 0005576细胞外区域80.0014DKK3, FRZB, DKK2, WIF1, SFRP4, SFRP2, SFRP1, DKK1

分子功能

Molecular function

MSTNGO: 0048179激活素受体复合体41.99E-08TGFBR1, ACVR2A, ACVR1B, ACVR2B
GO: 0048185激活素结合62.20E-13FST, TGFBR1, ACVR2A, ACVR1B, FSTL3, ACVR2B
BDNFGO: 0017002激活素受体活性44.83E-08TGFBR1, ACVR2A, ACVR1B, ACVR2B
DKK3GO: 0017147Wnt蛋白结合51.02E-08LRP5, FRZB, SFRP4, SFRP2, SFRP1
GO: 0005102信号受体结合60.0105DKK3, DKK2, WIF1, SFRP2, SFRP1, DKK1
GO: 0005515蛋白质结合100.021DKK3, LRP5, FRZB, DKK2, WIF1, SFRP4, CTNNB1, SFRP2, SFRP1, DKK1
GO: 0030545信号受体调节活性40.0462DKK3, DKK2, SFRP2, DKK1
GO:0017147Wnt蛋白结合51.02E-08LRP5, FRZB, SFRP4, SFRP2, SFRP1
生物学过程Biological processMSTNGO: 0032925激活素受体信号通路的调控56.04E-09FST, SMAD2, ACVR2A, ACVR1B, FSTL3
GO: 0007178跨膜受体蛋白丝氨酸/苏氨酸激酶信号通路77.99E-09FST, MSTN, SMAD2, TGFBR1, ACVR2A, ACVR1B,ACVR2B
GO: 0045595细胞分化调控107.99E-09MYOD1, FST, MYOG, MSTN, SMAD2, TGFBR1, ACVR2A, MYF5, ACVR1B, FSTL3
BDNFGO: 0038179神经营养因子信号通路54.78E-09BDNF, NTRK2, NTF4, NGF, SORT1
DKK3GO: 0090090典型Wnt信号通路的负调控94.90E-15DKK3, FRZB, DKK2, SFRP4, CTNNB1, KREMEN1, SFRP2, SFRP1, DKK1
GO: 0045597细胞分化的正调控74.31E-06LRP5, FRZB, WIF1, SFRP4, CTNNB1, SFRP2, SFRP1

图4

MSTN、BDNF、DKK3基因在信阳水牛成年牛11种组织中的相对表达情况注：不同小写字母表示差异显著（P<0.05），不同大写字母表示差异极显著（P<0.01）。"

图5

MSTN、BDNF在信阳水牛原代成肌细胞诱导分化过程中的相对表达"

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