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对鲑鱼虱(Lepeophtheirus salmonis (Krøyer, 1837))细胞色素 P450 基因的全基因组调查。

Genome-wide survey of cytochrome P450 genes in the salmon louse Lepeophtheirus salmonis (Krøyer, 1837).

机构信息

Institute of Aquaculture, University of Stirling, Stirling, FK9 4LA, Scotland, UK.

Department of Microbiology, University of Tennessee, Memphis, TN, 38163, USA.

出版信息

Parasit Vectors. 2019 Nov 27;12(1):563. doi: 10.1186/s13071-019-3808-x.

Abstract

BACKGROUND

The salmon louse (Lepeophtheirus salmonis) infests farmed and wild salmonid fishes, causing considerable economic damage to the salmon farming industry. Infestations of farmed salmon are controlled using a combination of non-medicinal approaches and veterinary drug treatments. While L. salmonis has developed resistance to most available salmon delousing agents, relatively little is known about the molecular mechanisms involved. Members of the cytochrome P450 (CYP) superfamily are typically monooxygenases, some of which are involved in the biosynthesis and metabolism of endogenous compounds, while others have central roles in the detoxification of xenobiotics. In terrestrial arthropods, insecticide resistance can be based on the enhanced expression of CYPs. The reported research aimed to characterise the CYP superfamily in L. salmonis and assess its potential roles in drug resistance.

METHODS

Lepeophtheirus salmonis CYPs were identified by homology searches of the genome and transcriptome of the parasite. CYP transcript abundance in drug susceptible and multi-resistant L. salmonis was assessed by quantitative reverse transcription PCR, taking into account both constitutive expression and expression in parasites exposed to sublethal levels of salmon delousing agents, ecdysteroids and environmental chemicals.

RESULTS

The above strategy led to the identification of 25 CYP genes/pseudogenes in L. salmonis, making its CYP superfamily the most compact characterised for any arthropod to date. Lepeophtheirus salmonis possesses homologues of a number of arthropod CYP genes with roles in ecdysteroid metabolism, such as the fruit fly genes disembodied, shadow, shade, spook and Cyp18a1. CYP transcript expression did not differ between one drug susceptible and one multi-resistant strain of L. salmonis. Exposure of L. salmonis to emamectin benzoate or deltamethrin caused the transcriptional upregulation of certain CYPs. In contrast, neither ecdysteroid nor benzo[a]pyrene exposure affected CYP transcription significantly.

CONCLUSIONS

The parasite L. salmonis is demonstrated to possess the most compact CYP superfamily characterised for any arthropod to date. The complement of CYP genes in L. salmonis includes conserved CYP genes involved in ecdysteroid biosynthesis and metabolism, as well as drug-inducible CYP genes. The present study does not provide evidence for a role of CYP genes in the decreased susceptibility of the multiresistant parasite strain studied.

摘要

背景

鲑虱(Lepeophtheirus salmonis)寄生在养殖和野生鲑鱼科鱼类上,给鲑鱼养殖业造成了相当大的经济损失。养殖鲑鱼的寄生虫感染通过非药物方法和兽医药物治疗相结合来控制。虽然鲑虱已经对大多数可用的鲑鱼除虱剂产生了抗药性,但对涉及的分子机制知之甚少。细胞色素 P450(CYP)超级家族的成员通常是单加氧酶,其中一些参与内源性化合物的生物合成和代谢,而另一些则在异生物质的解毒中起核心作用。在陆生节肢动物中,杀虫剂抗性可以基于 CYP 的增强表达。本研究旨在描述鲑虱的 CYP 超级家族,并评估其在药物抗性中的潜在作用。

方法

通过寄生虫基因组和转录组的同源搜索,鉴定鲑虱的 CYP。通过定量逆转录 PCR 评估对药物敏感和多耐药鲑虱的 CYP 转录丰度,同时考虑到暴露于亚致死水平的鲑鱼除虱剂、蜕皮激素和环境化学物质下寄生虫的组成型表达和表达。

结果

该策略导致在鲑虱中鉴定出 25 个 CYP 基因/假基因,使其 CYP 超级家族成为迄今为止描述的任何节肢动物中最紧凑的。鲑虱具有与蜕皮激素代谢有关的许多节肢动物 CYP 基因的同源物,如果蝇基因 disembodied、shadow、shade、spook 和 Cyp18a1。一种药物敏感和一种多耐药鲑虱品系之间的 CYP 转录表达没有差异。暴露于埃玛菌素苯甲酸酯或溴氰菊酯会导致某些 CYP 的转录上调。相比之下,蜕皮激素或苯并[a]芘暴露对 CYP 转录没有显著影响。

结论

本研究证明,寄生虫鲑虱拥有迄今为止描述的任何节肢动物中最紧凑的 CYP 超级家族。鲑虱的 CYP 基因包括参与蜕皮激素生物合成和代谢的保守 CYP 基因,以及药物诱导的 CYP 基因。本研究没有提供证据表明 CYP 基因在研究的多耐药寄生虫菌株的低敏感性中起作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6038/6880348/5ed50cf63698/13071_2019_3808_Fig1_HTML.jpg

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