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双斑青蟹(Charybdis bimaculata)血细胞中的抗菌肽基因及其表达特性分析

Effects of methyl farnesoate on Krüppel homolog 1 (Kr-h1) during vitellogenesis in the Chinese mitten crab (Eriocheir sinensis).

机构信息

College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China.

College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China.

出版信息

Anim Reprod Sci. 2021 Jan;224:106653. doi: 10.1016/j.anireprosci.2020.106653. Epub 2020 Nov 22.

DOI:10.1016/j.anireprosci.2020.106653
PMID:33249353
Abstract

Methyl farnesoate (MF), a de-epoxidized form of juvenile hormone (JH) Ⅲ in insects, may regulate developmental processes such as reproduction and ovarian maturation in crustaceans. Krüppel homolog 1 (Kr-h1) is a target response gene for the methoprene-tolerant (Met) protein that is a component of the JH signaling pathway in insects. In the present study, Es-Kr-h1 was cloned from E. sinensis and characterized to ascertain whether JH/MF signaling in insects is conserved in crustaceans. The findings with molecular structure analysis indicated Es-Kr-h1 contains seven zinc finger motifs (Zn2-Zn8) commonly conserved in other crustaceans, but the Zn1 motif was not detected to be present. The PCR results indicated that relative abundance of Es-Kr-h1 mRNA transcript in the hepatopancreas was greatest in the Stage Ⅱ, followed by the Stage Ⅳ ovarian developmental categories. The relative abundance of Es-Kr-h1 mRNA transcript in vitro was greater after MF addition to the hepatopancreas, however, not the ovarian tissues. The results from in vivo and eyestalk ablation experiments indicated the relative abundance of Es-Kr-h1 mRNA transcript was greater after MF treatment and bilateral eyestalk removal in the hepatopancreas, however, not ovarian tissues. Notably, there were effects of MF on relative abundance of Es-Kr-h1 mRNA transcript pattern. The Es-Kr-h1 protein, therefore, may be involved in MF-mediated vitellogenesis resulting from the response to Es-Met in E. sinensis, and the JH/MF signaling pathway is potentially conserved in crustaceans.

摘要

双羟茉莉酸甲酯(MF),一种昆虫保幼激素(JH)Ⅲ的去环氧化形式,可能调节甲壳动物的生殖和卵巢成熟等发育过程。Krüppel 同源物 1(Kr-h1)是蜕皮激素耐受(Met)蛋白的靶基因响应,Met 蛋白是昆虫 JH 信号通路的组成部分。在本研究中,从中华绒螯蟹中克隆了 Es-Kr-h1,并对其进行了特征描述,以确定昆虫中的 JH/MF 信号是否在甲壳动物中保守。分子结构分析的结果表明,Es-Kr-h1 含有七个锌指结构域(Zn2-Zn8),这在其他甲壳动物中普遍保守,但未检测到存在 Zn1 结构域。PCR 结果表明,Es-Kr-h1 mRNA 转录本在肝胰腺中的相对丰度在 Stage Ⅱ最大,其次是 Stage Ⅳ卵巢发育类别。MF 加入肝胰腺后,Es-Kr-h1 mRNA 转录本的相对丰度增加,但卵巢组织则不然。体内和眼柄切除实验的结果表明,MF 处理和双侧眼柄切除后,Es-Kr-h1 mRNA 转录本在肝胰腺中的相对丰度增加,但卵巢组织则不然。值得注意的是,MF 对 Es-Kr-h1 mRNA 转录本模式的相对丰度有影响。因此,Es-Kr-h1 蛋白可能参与 MF 介导的卵黄发生,这是中华绒螯蟹对 Es-Met 反应的结果,并且 JH/MF 信号通路在甲壳动物中可能是保守的。

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