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全球毒蝎 Centruroides noxius 的转录组分析:从一个古老的蝎种中发现新的毒素家族和进化见解。

Global transcriptome analysis of the scorpion Centruroides noxius: new toxin families and evolutionary insights from an ancestral scorpion species.

机构信息

Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados, Irapuato, Guanajuato, México.

出版信息

PLoS One. 2012;7(8):e43331. doi: 10.1371/journal.pone.0043331. Epub 2012 Aug 17.

Abstract

Scorpion venoms have been studied for decades, leading to the identification of hundreds of different toxins with medical and pharmacological implications. However, little emphasis has been given to the description of these arthropods from cellular and evolutionary perspectives. In this report, we describe a transcriptomic analysis of the Mexican scorpion Centruroides noxius Hoffmann, performed with a pyrosequencing platform. Three independent sequencing experiments were carried out, each including three different cDNA libraries constructed from RNA extracted from the whole body of the scorpion after telson removal, and from the venom gland before and after venom extraction. Over three million reads were obtained and assembled in almost 19000 isogroups. Within the telson-specific sequences, 72 isogroups (0.4% of total unique transcripts) were found to be similar to toxins previously reported in other scorpion species, spiders and sea anemones. The annotation pipeline also revealed the presence of important elements of the small non-coding RNA processing machinery, as well as microRNA candidates. A phylogenomic analysis of concatenated essential genes evidenced differential evolution rates in this species, particularly in ribosomal proteins and proteasome components. Additionally, statistical comparison of transcript abundance before and after venom extraction showed that 3% and 2% of the assembled isogroups had higher expression levels in the active and replenishing gland, respectively. Thus, our sequencing and annotation strategies provide a general view of the cellular and molecular processes that take place in these arthropods, allowed the discovery of new pharmacological and biotechnological targets and uncovered several regulatory and metabolic responses behind the assembly of the scorpion venom. The results obtained in this report represent the first high-throughput study that thoroughly describes the universe of genes that are expressed in the scorpion Centruroides noxius Hoffmann, a highly relevant organism from medical and evolutionary perspectives.

摘要

蝎毒液已经研究了几十年,导致了数百种不同毒素的鉴定,这些毒素具有医学和药理学意义。然而,从细胞和进化的角度来描述这些节肢动物的研究却很少受到重视。在本报告中,我们描述了对墨西哥蝎子 Centruroides noxius Hoffmann 的转录组分析,该分析是使用焦磷酸测序平台进行的。进行了三个独立的测序实验,每个实验都包括从去除尾节后的蝎子整体和毒液腺在提取毒液前后提取的 RNA 构建的三个不同 cDNA 文库。获得了超过三百万个读数,并在近 19000 个同工型中组装。在尾节特异性序列中,发现 72 个同工型(总独特转录本的 0.4%)与先前在其他蝎子、蜘蛛和海葵中报道的毒素相似。注释管道还揭示了小非编码 RNA 加工机制的重要元件以及 microRNA 候选物的存在。串联必需基因的系统基因组分析证明了该物种的进化速率存在差异,特别是在核糖体蛋白和蛋白酶体成分中。此外,对毒液提取前后转录本丰度的统计比较表明,在活性和补充腺中,分别有 3%和 2%的组装同工型具有更高的表达水平。因此,我们的测序和注释策略提供了这些节肢动物发生的细胞和分子过程的总体视图,发现了新的药理学和生物技术靶标,并揭示了在蝎子毒液组装背后的几个调节和代谢反应。本报告中的结果代表了首次高通量研究,彻底描述了在医学和进化角度上具有重要意义的 Centruroides noxius Hoffmann 蝎子中表达的基因宇宙。

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