浙江农业学报 ›› 2025, Vol. 37 ›› Issue (5): 1029-1044.DOI: 10.3969/j.issn.1004-1524.20240463

• 动物科学 • 上一篇    下一篇

基于串联质量标签(TMT)技术的宁都黄鸡大小睾丸蛋白质组学研究

马荆鄂(), 吴斯琪, 万淑敏, 熊信威, 许继国, 许晶, 饶友生, 周敏()   

  1. 南昌师范学院 生物技术研究院,家禽遗传改良江西省重点实验室,江西 南昌 330032
  • 收稿日期:2024-05-27 出版日期:2025-05-25 发布日期:2025-06-11
  • 作者简介:马荆鄂(1987—),女,湖北荆州人,博士,副教授,研究方向为家禽遗传育种与繁殖。E-mail:majinge@ncnu.edu.cn
  • 通讯作者: *周敏,E-mail:623808803@qq.com
  • 基金资助:
    国家自然科学基金(32160784);江西省高等学校教学改革研究重点课题(JXJG-23-23-5)

Proteomic study of large and small testes in Ningdu Yellow chicken based on tandem mass tags(TMT) technology

MA Jing'e(), WU Siqi, WAN Shumin, XIONG Xinwei, XU Jiguo, XU Jing, RAO Yousheng, ZHOU Min()   

  1. Jiangxi Provincial Key Laboratory of Poultry Genetic Improvement, Institution of Biological Technology, Nanchang Normal University, Nanchang 330032, China
  • Received:2024-05-27 Online:2025-05-25 Published:2025-06-11

摘要: 试验旨在探讨成年宁都黄鸡睾丸的蛋白质表达谱,筛选大小睾丸个体差异表达蛋白和功能通路,为繁殖性能选育提供基础数据。以6只22周龄宁都黄鸡的睾丸组织为研究对象,分为大睾丸组(H-TES)和小睾丸组(L-TES),采用串联质量标签(tandem mass tags, TMT)技术对2组个体睾丸蛋白质进行差异表达、功能富集和基因互作分析,初步筛选候选蛋白质,分析这些蛋白质在6个个体的表达量与6个睾丸性状指标(左侧睾丸重、右侧睾丸重、总睾丸重、左侧睾丸指数、右侧睾丸指数、总睾丸指数)之间的相关性,分析候选蛋白质的功能。结果表明,宁都黄鸡睾丸中共鉴定到4 683个蛋白质,筛选到144个差异表达蛋白质。GO分析结果表明,差异蛋白质主要参与精子结构组成,具有尿蛋白-谷氨酸连接酶活性、胸腺嘧啶结合、脱氢抗坏血酸跨膜转运体活性等功能。差异蛋白质显著富集的KEGG通路为精氨酸和脯氨酸代谢、泛酸和辅酶A的生物合成、β-丙氨酸代谢等。蛋白质互作网络中有34个差异蛋白质,初步筛选出30个候选蛋白质。结合相关性分析和文献查阅,筛选出16个关键候选蛋白质,即SPAG6、TPPP2、ENKUR、ODF2、SAXO1、TCP11、DNALI1、CCDC63、TSSK3、TEKT2、TEKT3、ALDH2、DNAH5、CCDC40、CCDC39、DNAH10。结合关键候选蛋白质的功能,初步确定精氨酸和脯氨酸代谢、泛酸和辅酶A生物合成、β-丙氨酸代谢、紧密连接、间隙连接是睾丸性状调控关键KEGG通路。研究结果为地方鸡种睾丸质量性状选育提供了基础依据。

关键词: 宁都黄鸡, 睾丸, TMT, 蛋白质组学, 候选基因

Abstract:

This experiment aimed to investigate the protein expression profiles in testes of adult Ningdu Huang chickens, screen differentially expressed proteins and functional pathways between individuals with large and small testes, and provide foundational data for breeding selection of reproductive performance. Using testicular tissues from six 22-week-old Ningdu Huang chickens divided into large-testis(H-TES) group and small-testis(L-TES) group, tandem mass tags(TMT) technology was employed to conduct differential expression analysis, functional enrichment analysis, and gene interaction analysis of testicular proteins between the two groups. Candidate proteins were preliminarily screened, and correlations between the expression levels of these proteins in six individuals and six testicular trait indicators (left testis weight, right testis weight, total testis weight, left testis index, right testis index, total testis index) were analyzed to investigate protein functions. Results showed that 4 683 proteins were identified in Ningdu Huang chicken testes, with 144 differentially expressed proteins screened. GO analysis revealed that these differential expressed proteins primarily participate in sperm structural composition and possess functions including ubulin-glutamic acid ligase activity, thymine binding, and dehydroascorbic acid transmembrane transporter activity, etc. Significantly enriched KEGG pathways included arginine and proline metabolism, pantothenate and CoA biosynthesis, and β-alanine metabolism, etc. The protein interaction network contained 34 differential proteins, and 30 candidate proteins were preliminarily identified. Through correlation analysis and literature review, 16 key candidate proteins were screened: SPAG6, TPPP2, ENKUR, ODF2, SAXO1, TCP11, DNALI1, CCDC63, TSSK3, TEKT2, TEKT3, ALDH2, DNAH5, CCDC40, CCDC39, and DNAH10. Based on their functions, key regulatory KEGG pathways for testicular traits were identified as arginine and proline metabolism, pantothenate and CoA biosynthesis, β-alanine metabolism, tight junction, and gap junction. These findings provide fundamental data for breeding selection of testicular quality traits in local chicken breeds.

Key words: Ningdu yellow chicken, testis, tandem mass tag(TMT), proteome, candidate gene

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