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来自海虱(Caligus rogercresseyi)的微小RNA生物合成途径:在灭虱药物反应中的新作用。

MicroRNA biogenesis pathway from the salmon louse (Caligus rogercresseyi): emerging role in delousing drug response.

作者信息

Valenzuela-Miranda Diego, Nuñez-Acuña Gustavo, Valenzuela-Muñoz Valentina, Asgari Sassan, Gallardo-Escárate Cristian

机构信息

Laboratory of Biotechnology and Aquatic Genomics, Interdisciplinary Center for Aquaculture Research (INCAR), University of Concepción, P. O. Box 160-C, Concepción, Chile.

Australian Infectious Disease Research Centre, School of Biological Sciences, The University of Queensland, Brisbane, QLD, Australia.

出版信息

Gene. 2015 Jan 25;555(2):231-41. doi: 10.1016/j.gene.2014.11.008. Epub 2014 Nov 7.

Abstract

Despite the increasing evidence of the importance of microRNAs (miRNAs) in the regulation of multiple biological processes, the molecular bases supporting this regulation are still barely understood in crustaceans. Therefore, the molecular characterization and transcriptome modulation of the miRNA biogenesis pathway were evaluated in the salmon louse Caligus rogercresseyi, an ectoparasite that constitutes one of the biggest concerns for salmonid aquaculture industry. Hence, RNA-Seq analysis was conducted from six different developmental stages, and also after bioassays with delousing drugs Deltamethrin and Azamethiphos using adult individuals. In silico analysis evidenced 24 putative genes involved in the miRNA pathway such as biogenesis, transport, maturation and miRNA-target interaction. Moreover, 243 putative single nucleotide polymorphisms (SNPs) were identified, 15 of which showed non-synonym mutations. RNA-Seq analysis revealed that CCR4-Not complex subunit 3 (CNOT3) was upregulated at earlier developmental stages (nauplius I-II and copepodid), and also after the exposure to Azamethiphos, but not to Deltamethrin. In contrast, the subunit 7 (CNOT7) showed an inverse expression pattern. Different Argonaute transcripts were associated to chalimus and adult stages, revealing specific expression patterns in response to antiparasitic drugs. Our results suggest novel insights into the regulatory network of the post-transcriptional gene regulation in C. rogercresseyi mediated by miRNAs, evidencing a putative role during the ontogeny and drug response.

摘要

尽管越来越多的证据表明微小RNA(miRNA)在多种生物过程的调节中具有重要作用,但在甲壳类动物中,支持这种调节的分子基础仍几乎不为人知。因此,在鲑鱼虱Caligus rogercresseyi中评估了miRNA生物合成途径的分子特征和转录组调控,鲑鱼虱是一种体外寄生虫,是鲑鱼养殖业最关注的问题之一。因此,对六个不同发育阶段进行了RNA测序分析,并且在使用溴氰菊酯和谷硫磷这两种灭虱药物对成年个体进行生物测定后也进行了RNA测序分析。计算机分析证明有24个推定基因参与miRNA途径,如生物合成、转运、成熟和miRNA-靶标相互作用。此外,鉴定出243个推定的单核苷酸多态性(SNP),其中15个显示非同义突变。RNA测序分析表明,CCR4-Not复合物亚基3(CNOT3)在发育早期阶段(无节幼虫I-II期和桡足幼体期)以及暴露于谷硫磷后上调,但在暴露于溴氰菊酯后未上调。相比之下,亚基7(CNOT7)表现出相反的表达模式。不同的AGO蛋白转录本与夏卵幼虫期和成虫期相关,揭示了对抗寄生虫药物的特异性表达模式。我们的结果为miRNA介导的C. rogercresseyi转录后基因调控网络提供了新的见解,证明了其在个体发育和药物反应中的假定作用。

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