细粒棘球绦虫原头蚴蛋白表达谱分析

doi:10.11843/j.issn.0366-6964.2017.08.017

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (8): 1519-1528.doi: 10.11843/j.issn.0366-6964.2017.08.017

细粒棘球绦虫原头蚴蛋白表达谱分析

王正荣¹, 薄新文^1*, 张艳艳¹, 马勋^1,2, 王智欣^1,3, 卢平萍^1,2

1. 省部共建绵羊遗传改良与健康养殖国家重点实验室/新疆农垦科学院畜牧兽医研究所, 石河子 832000;
2. 石河子大学动物科技学院, 石河子 832000;
3. 中国农业大学动物医学院, 北京 100193

收稿日期:2017-03-24 出版日期:2017-08-23 发布日期:2017-08-12
通讯作者: 薄新文,男,研究员,主要从事寄生虫分子生物学研究,E-mail:boxinwen@yahoo.com.cn
作者简介:王正荣(1984-),男,甘肃定西人,硕士,助理研究员,主要从事寄生虫分子生物学研究,E-mail:wzrtiger@sina.com
基金资助:
国家重点基础研究发展计划（973计划）项目（2015CB150300）；国家自然科学基金（31360608）；新疆生产建设兵团科技攻关项目（2014BA002）；新疆生产建设兵团国际科技合作项目（2013BC001）；新疆生产建设兵团绵羊繁育生物技术重点实验室开放课题（2013KLS06）

Protein Profile Analyses of Protoscoleces in Echinococcus granulosus

WANG Zheng-rong¹, BO Xin-wen^1*, ZHANG Yan-yan¹, MA Xun^1,2, WANG Zhi-xin^1,3, LU Ping-ping^1,2

1. State Key Labaratory of Sheep Genetic Improvement and Healthy Production/Institute of Animal Husbandry and Veterinary, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832000, China;
2. College of Animal Science and Technology, Shihezi University, Shihezi 832000, China;
3. College of Veterinary Medicine, China Agricultural University, Beijing 100193, China

Received:2017-03-24 Online:2017-08-23 Published:2017-08-12

摘要/Abstract

摘要：

旨在探索细粒棘球绦虫原头蚴蛋白表达特性，揭示在此发育阶段虫体主要的功能蛋白以及信号通路，为揭示虫体的发育调控、筛选疫苗以及药物靶标奠定基础。本试验采用LC-MS/MS技术对虫体原头蚴表达的蛋白质进行研究，然后对获得数据进行GO、KOG以及KEGG等分析；最后采用qRT-PCR和原位杂交技术对鉴定得到的Wnt信号通路的关键基因β-catenin进行了差异表达和组织定位研究。结果显示，本研究共鉴定得到7 172个肽段，1 197个蛋白质，其中包括多个潜在的抗原蛋白质，如四跨膜蛋白超家族、6-磷酸葡萄糖酸脱氢酶、表皮抗原蛋白和烯醇酶等。信号通路分析结果显示其中632个蛋白参与276个调控通路，诸如与虫体发育紧密相关的Wnt、Notch、Hedgehog、NF-κB、cAMP以及胆酸盐信号通路。对β-catenin的验证结果显示，此基因分布于细粒棘球绦虫原头蚴顶突的小刺以及散在的细胞中，同时其相对转录量在体外培养0~10 d的过程中呈递减趋势。综上，本研究解析了细粒棘球绦虫原头蚴的蛋白质表达元件，揭示了原头蚴主要的调控信号通路，为虫体的生长发育机制以及包虫病的有效防控提供了理论依据。

关键词: 细粒棘球绦虫, 原头蚴, 蛋白, 表达谱, 信号通路

Abstract:

The aim of this study was to explore the protein expression characteristics of protoscoleces, to reveal the main functional proteins and signaling pathways during the development of protoscoleces in Echinococcus granulusos, to lay the foundation for the mechanisms of development regulation of protoscoleces and screening target genes for vaccines and drugs. The LC-MS/MS technology was used to analyze the proteins expressed in the protoscoleces, then the proteins were analyzed based on GO, KOG and KEGG. Finally, the qRT-PCR and in situ hybridization were used to analyze the different expression and localization of the β-catenin which is the key gene of the Wnt signaling pathway. The results showed that the present study identified 7 172 peptides, 1 197 proteins, including multiple potential antigenic proteins, such as tetraspanin superfamily, 6-phosphogluconate dehydrogenase, enolase and epidermal antigenic proteins. Signal pathway analysis showed that 632 of these proteins involved in 276 pathways, such as Wnt, Notch, Hedgehog, NF-κB, cAMP and bile acid signaling pathways closely related to development of E. granulosus. The in situ hybridization analysis showed that the β-catenin was mainly distributed in the hooks and dispersed cells of the protoscoleces. Meanwhile, the relative transcription of β-catenin was gradually decreased during 0 to 10 days in vitro culture. In summary, this study analyzed the protein expression elements of protoscoleces, revealed the main regulatory pathways of protoscolex development, and provided a theoretical basis for the growth mechanisms of protoscoleces and effective prevention of hydatid disease.

Key words: Echinococcus granulosus, protoscolex, protein, profile, signaling pathway

中图分类号:

S852.734

王正荣, 薄新文, 张艳艳, 马勋, 王智欣, 卢平萍. 细粒棘球绦虫原头蚴蛋白表达谱分析[J]. 畜牧兽医学报, 2017, 48(8): 1519-1528.

WANG Zheng-rong, BO Xin-wen, ZHANG Yan-yan, MA Xun, WANG Zhi-xin, LU Ping-ping. Protein Profile Analyses of Protoscoleces in Echinococcus granulosus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(8): 1519-1528.

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细粒棘球绦虫原头蚴蛋白表达谱分析

Protein Profile Analyses of Protoscoleces in Echinococcus granulosus

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