华北农学报 ›› 2025, Vol. 40 ›› Issue (3): 182-189. doi: 10.7668/hbnxb.20195468

所属专题: 畜牧

• 畜牧·水产·兽医 • 上一篇    下一篇

甘加型藏羊TACR3基因克隆及在发情周期和乏情期输卵管、子宫中的表达

赵国勇1, 高何璇2, 刘晓洁1, 何玉琴1, , 陶乐凯1, 蔡永强1, 张光敏1   

  1. 1 甘肃农业大学 生命科学技术学院,甘肃 兰州 730070
    2 兰州银行网络金融部,甘肃 兰州 730070
  • 收稿日期:2024-09-30 出版日期:2025-06-28
  • 通讯作者:
    何玉琴(1963—),女,甘肃兰州人,教授,博士,硕士生导师,主要从事动物生理学研究。
  • 作者简介:

    赵国勇(1997—),男,甘肃武威人,在读硕士,主要从事动物生理学、生物化学与分子生物学研究。

  • 基金资助:
    国家自然科学基金项目(31960689)

Cloning of TACR3 Gene and Its Expression in Oviduct and Uterus During Estrous Cycle and Anestrus of Ganjia Tibetan Sheep

ZHAO Guoyong1, GAO Hexuan2, LIU Xiaojie1, HE Yuqin1, , TAO Lekai1, CAI Yongqiang1, ZHANG Guangmin1   

  1. 1 College of Life Science and Technology,Gansu Agricultural University,Lanzhou 730070,China
    2 Network Finance Department,Bank of Lanzhou,Lanzhou 730070,China
  • Received:2024-09-30 Published:2025-06-28

摘要:

为探究速激肽3受体基因(TACR3)功能及其编码产物神经激肽B受体(NK3R)在甘加型藏羊输卵管和子宫中的表达与分布,以发情周期和乏情期甘加型藏羊下丘脑、输卵管和子宫组织为样本,通过巢式PCR反应克隆TACR3基因编码区序列(CDS),利用STRING蛋白质互作数据库、基因本体论(GO)和京都基因与基因组百科全书(KEGG)预测分析TACR3互作蛋白的生物学功能;通过实时荧光定量PCR(qRT-PCR)、蛋白免疫印迹(WB)和免疫组织化学(IHC)方法分析TACR3基因及其编码蛋白NK3R在甘加型藏羊发情周期和乏情期输卵管和子宫的分布及表达规律。结果显示,甘加型藏羊TACR3基因开放阅读框全长1 95 bp,编码464个氨基酸,未发现信号肽结构,存在7次跨膜结构;TACR3可能参与神经肽信号通路、GnRH信号通路和钙信号通路等生物学过程。TACR3编码的NK3R蛋白主要表达在输卵管的黏膜上皮、纤毛细胞、分泌细胞和子宫的子宫腺、基质细胞中。输卵管中TACR3 mRNA在发情后期表达量最高,NK3R蛋白在发情期表达量最高;子宫中TACR3 mRNA和蛋白均在发情后期表达量最高。结果表明,TACR3基因在动物进化中较为保守,不同发情时期的输卵管和子宫组织均有TACR3 mRNA及蛋白表达,其可能在甘加型藏羊生殖过程中发挥重要作用。

关键词: 甘加型藏羊, 速激肽3受体基因(TACR3), 神经激肽B受体(NK3R), 基因克隆, 发情周期

Abstract:

To investigate the function of tachykinin 3 receptor gene(TACR3)and the expression and distribution of its encoded product neurokinin B receptor(NK3R)in the oviduct and uterus of Ganjia Tibetan sheep,hypothalamus,oviduct and uterus tissues during estrous cycle and anestrus of Ganjia Tibetan sheep were taken as samples.The TACR3 gene coding sequence(CDS)was cloned by nested PCR.The biological function of TACR3 interacting proteins was predicted using STRING protein interaction database,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG).Real-time Quantitative PCR(qRT-PCR),Western Blot and immunohistochemistry(IHC)were used to analyze the distribution and expression of TACR3 gene and NK3R in the oviduct and uterus during estrous cycle and anestrus of Ganjia Tibetan sheep.The results showed that the total length of the open reading frame of TACR3 gene in Ganjia Tibetan sheep was 1 395 bp,encoding 464 amino acids,and no signal peptide structure was found,with seven transmembrane structures.TACR3 may participate in biological processes such as neuropeptide signaling pathway,GnRH signaling pathway and calcium signaling pathway.The NK3R protein encoded by TACR3 was mainly expressed in the mucosal epithelium,ciliary cells,secretory cells of oviduct,uterine glands and stromal cells of uterus.In oviduct,the expression level of TACR3 mRNA was the highest in the metestrus,and the expression level of NK3R protein was the highest in the estrus.The expression levels of TACR3 mRNA and protein in uterus were the highest in the metestrus.The results showed that TACR3 was conserved in animal evolution,and TACR3 mRNA and protein were expressed in oviduct and uterus tissues at different estrus periods,which may play an important role in the reproductive process of Ganjia Tibetan sheep.

Key words: Ganjia Tibetan sheep, Tachykinin receptor 3 gene(TACR3), Neurokinin B receptor(NK3R), Gene cloning, Estrous cycle

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引用本文

赵国勇, 高何璇, 刘晓洁, 何玉琴, 陶乐凯, 蔡永强, 张光敏. 甘加型藏羊TACR3基因克隆及在发情周期和乏情期输卵管、子宫中的表达[J]. 华北农学报, 2025, 40(3): 182-189. doi: 10.7668/hbnxb.20195468.

ZHAO Guoyong, GAO Hexuan, LIU Xiaojie, HE Yuqin, TAO Lekai, CAI Yongqiang, ZHANG Guangmin. Cloning of TACR3 Gene and Its Expression in Oviduct and Uterus During Estrous Cycle and Anestrus of Ganjia Tibetan Sheep[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(3): 182-189. doi: 10.7668/hbnxb.20195468.