Transcriptomic analysis of crustacean neuropeptide signaling during the moult cycle in the green shore crab, Carcinus maenas.
Transcriptomic analysis of crustacean neuropeptide signaling during the moult cycle in the green shore crab, Carcinus maenas.
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DOI：
10.1186/s12864-018-5057-3
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发表时间：
2018-09-26
期刊：
影响因子：
4.4
通讯作者：
Wilcockson DC
中科院分区：
文献类型：
--
作者：
Oliphant A;Alexander JL;Swain MT;Webster SG;Wilcockson DC
蜕皮是所有节肢动物进行外骨骼蜕皮的一种先天行为。在昆虫中，相互作用的神经肽的复杂套件编排蜕皮得到了很好的研究，但甲壳动物蜕皮盒的细节是支离破碎的，我们对这个过程的理解是相对粗糙的，防止有意义的进化比较。为了开始解决这个问题，我们确定了转录编码的神经肽和它们的假定受体在中枢神经系统（CNS）和Y-器官（YO）内的螃蟹，巨蟹，并绘制了他们的表达谱准确定义的蜕皮周期使用RNA测序的阶段。我们还研究了表皮衍生的YO内的基因表达，唯一确定的作用是蜕皮激素蜕皮激素的合成，以阐明可能在蜕皮中具有功能的肽和G蛋白偶联受体（GPCR）。CNS转录组的转录组挖掘产生了代表47个神经肽家族和66个推定的GPCR的神经肽转录本。在整个蜕皮周期中差异表达的神经肽转录物包括carcikinin，甲壳动物高血糖激素-2，甲壳动物心脏活性肽，而一个单一的假定的神经肽受体，proctolin R1，差异表达。特别是Carcikinin mRNA表现出显着增加，在蜕皮前的表达，这表明在蜕皮调节的作用。甲壳动物高血糖激素-2 mRNA的表达升高后和蜕皮前，而甲壳动物心脏活性肽，调节昆虫蜕皮和甲壳动物蜕皮期间的定型运动活动中发挥作用，在蜕皮前升高。在YO中，几种假定的神经肽受体转录本在整个换羽周期中差异表达，神经肽neuroparsin-1的mRNA也是如此。虽然预期推定的神经肽受体的差异基因表达，但神经肽转录物的发现和差异表达是令人惊讶的。YO和表皮之间的GPCR转录本表达的分析显示11在YO中上调，因此现在是蜕皮肽控制的候选者。所提供的数据代表了推导的C. maenas神经肽组和推定的GPCR。重要的是，我们已经描述了这些转录本的差异表达谱，这些转录本在蜕皮程序的关键组织中准确地分期换羽周期。本研究为今后探索甲壳动物受体-配体对的功能提供了重要途径。本文的在线版本（10.1186/s12864-018-5057-3）包含补充材料，可供授权用户使用。
Ecdysis is an innate behaviour programme by which all arthropods moult their exoskeletons. The complex suite of interacting neuropeptides that orchestrate ecdysis is well studied in insects, but details of the crustacean ecdysis cassette are fragmented and our understanding of this process is comparatively crude, preventing a meaningful evolutionary comparison. To begin to address this issue we identified transcripts coding for neuropeptides and their putative receptors in the central nervous system (CNS) and Y-organs (YO) within the crab, Carcinus maenas, and mapped their expression profiles across accurately defined stages of the moult cycle using RNA-sequencing. We also studied gene expression within the epidermally-derived YO, the only defined role for which is the synthesis of ecdysteroid moulting hormones, to elucidate peptides and G protein-coupled receptors (GPCRs) that might have a function in ecdysis. Transcriptome mining of the CNS transcriptome yielded neuropeptide transcripts representing 47 neuropeptide families and 66 putative GPCRs. Neuropeptide transcripts that were differentially expressed across the moult cycle included carcikinin, crustacean hyperglycemic hormone-2, and crustacean cardioactive peptide, whilst a single putative neuropeptide receptor, proctolin R1, was differentially expressed. Carcikinin mRNA in particular exhibited dramatic increases in expression pre-moult, suggesting a role in ecdysis regulation. Crustacean hyperglycemic hormone-2 mRNA expression was elevated post- and pre-moult whilst that for crustacean cardioactive peptide, which regulates insect ecdysis and plays a role in stereotyped motor activity during crustacean ecdysis, was elevated in pre-moult. In the YO, several putative neuropeptide receptor transcripts were differentially expressed across the moult cycle, as was the mRNA for the neuropeptide, neuroparsin-1. Whilst differential gene expression of putative neuropeptide receptors was expected, the discovery and differential expression of neuropeptide transcripts was surprising. Analysis of GPCR transcript expression between YO and epidermis revealed 11 to be upregulated in the YO and thus are now candidates for peptide control of ecdysis. The data presented represent a comprehensive survey of the deduced C. maenas neuropeptidome and putative GPCRs. Importantly, we have described the differential expression profiles of these transcripts across accurately staged moult cycles in tissues key to the ecdysis programme. This study provides important avenues for the future exploration of functionality of receptor-ligand pairs in crustaceans. The online version of this article (10.1186/s12864-018-5057-3) contains supplementary material, which is available to authorized users.
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