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通过RNA干扰使瓦萨基因沉默会影响海虱(Caligus rogercresseyi)的胚胎发育和繁殖产出。

Silencing of the Vasa gene by RNA Interference Affects Embryonic Development and Reproductive Output in the Sea Louse Caligus rogercresseyi.

作者信息

Bustos Paulina, Schmitt Paulina, Brown Donald I, Farlora Rodolfo

机构信息

Laboratorio de Biotecnología Acuática y Genómica Reproductiva (LABYGER), Instituto de Biología, Facultad de Ciencias, Universidad de Valparaíso, Gran Bretaña 1111, 2360102, Valparaíso, Chile.

Doctorado en Acuicultura, Programa Cooperativo Universidad de Chile, Universidad Católica del Norte, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile.

出版信息

Mar Biotechnol (NY). 2023 Aug;25(4):612-623. doi: 10.1007/s10126-023-10232-5. Epub 2023 Aug 1.

Abstract

The sea louse Caligus rogercresseyi is a major ectoparasitic copepod that causes significant economic losses in the salmon farming industry. Despite recent advancements, the mechanisms underlying germline and embryo development in this species remain poorly understood. The Vasa gene encodes a highly conserved DEAD box helicase that is required for germ cell formation and function in many species. In this study, the Vasa gene was characterized in C. rogercresseyi, and its expression and function were analyzed. Phylogenetic analysis showed that the Cr-Vasa gene product formed clusters in clades with Vasa proteins from closely related species of crustaceans. Cr-Vasa gene expression patterns were assessed by qPCR, and the results showed a significantly higher relative expression level in adult females compared to copepodid, chalimus, and adult male stages. Tissue-specific localization of Cr-Vasa mRNA in C. rogercresseyi was determined using chromogenic in situ hybridization, and strong positive signal was observed in male testes, but also in the intestine and cuticle, while in females, it was observed in the ovaries, oocytes, cuticle, intestine, and egg strings. RNAi-mediated gene silencing of Cr-Vasa impacted embryonic development and reproductive output in adult female lice. Females from the dsVasa-treated group displayed unusual phenotypes, including shorter egg strings with numerous extra-embryonic inclusions, irregularly shaped abnormal embryos, and aborted egg strings. This study provides insights into the role of the Vasa gene in C. rogercresseyi embryonic development and reproductive output, which may have implications for the control of this parasitic copepod in the salmon farming industry.

摘要

海虱Caligus rogercresseyi是一种主要的体外寄生桡足类动物,给鲑鱼养殖业造成了巨大的经济损失。尽管最近有了进展,但该物种生殖细胞和胚胎发育的潜在机制仍知之甚少。Vasa基因编码一种高度保守的DEAD盒解旋酶,在许多物种中,它是生殖细胞形成和功能所必需的。在本研究中,对C. rogercresseyi中的Vasa基因进行了表征,并分析了其表达和功能。系统发育分析表明,Cr-Vasa基因产物与来自密切相关甲壳类物种的Vasa蛋白在进化枝中形成聚类。通过qPCR评估Cr-Vasa基因的表达模式,结果显示,与桡足幼体、无节幼体和成体雄性阶段相比,成年雌性中的相对表达水平显著更高。使用显色原位杂交确定了Cr-Vasa mRNA在C. rogercresseyi中的组织特异性定位,在雄性睾丸中观察到强阳性信号,在肠道和表皮中也观察到强阳性信号,而在雌性中,在卵巢、卵母细胞、表皮、肠道和卵串中观察到强阳性信号。RNAi介导的Cr-Vasa基因沉默影响成年雌性海虱的胚胎发育和繁殖产出。来自dsVasa处理组的雌性表现出异常表型, 包括带有大量胚外内含物的较短卵串、形状不规则的异常胚胎和流产的卵串。本研究深入了解了Vasa基因在C. rogercresseyi胚胎发育和繁殖产出中的作用,这可能对鲑鱼养殖业中这种寄生桡足类动物的控制具有重要意义。

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