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Transcriptomic Analysis of Neuropeptides and Peptide Hormones in the Barnacle Balanus amphitrite: Evidence of Roles in Larval Settlement

机译：在藤壶纹藤壶神经肽和肽类激素的转录组分析：在幼虫沉积作用的证据

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The barnacle Balanus amphitrite is a globally distributed marine crustacean and has been used as a model species for intertidal ecology and biofouling studies. Its life cycle consists of seven planktonic larval stages followed by a sessile juvenile/adult stage. The transitional processes between larval stages and juveniles are crucial for barnacle development and recruitment. Although some studies have been conducted on the neuroanatomy and neuroactive substances of the barnacle, a comprehensive understanding of neuropeptides and peptide hormones remains lacking. To better characterize barnacle neuropeptidome and its potential roles in larval settlement, an in silico identification of putative transcripts encoding neuropeptides/peptide hormones was performed, based on transcriptome of the barnacle B. amphitrite that has been recently sequenced. Potential cleavage sites andstructure of mature peptides were predicted through homology search of known arthropod peptides. In total, 16 neuropeptide families/subfamilies were predicted from the barnacle transcriptome, and 14 of them were confirmed as genuine neuropeptides by Rapid Amplification of cDNA Ends. Analysis of peptide precursor structures and mature sequences showed that some neuropeptides of B. amphitrite are novel isoforms and shared similar characteristics with their homologs from insects. The expression profiling of predicted neuropeptide genes revealed that pigment dispersing hormone, SIFamide, calcitonin, and B-type allatostatin had the highest expression level in cypris stage, while tachykinin-related peptide was down regulated in both cyprids and juveniles. Furthermore, an inhibitor of proprotein convertase related to peptide maturation effectively delayed larval metamorphosis. Combination of real-time PCR results and bioassay indicated that certain neuropeptides may play an important role in cypris settlement. Overall, new insight into neuropeptides/peptide hormones characterized in this study shall provide a platform for unraveling peptidergic control of barnacle larval behavior and settlement process.
机译：藤壶Balanus amphitrite是全球分布的海洋甲壳类动物，已被用作潮间生态学和生物污染研究的模型物种。它的生命周期包括七个浮游幼虫阶段，然后是无柄的幼体/成虫阶段。幼虫阶段和少年之间的过渡过程对于藤壶的发育和募集至关重要。尽管已经对藤壶的神经解剖学和神经活性物质进行了一些研究，但仍缺乏对神经肽和肽激素的全面了解。为了更好地表征藤壶神经肽组及其在幼虫沉降中的潜在作用，根据最近测序的藤壶双歧杆菌的转录组，对编码神经肽/肽激素的假定转录物进行了计算机鉴定。通过已知节肢动物肽的同源性搜索，可以预测成熟肽的潜在切割位点和结构。藤壶转录组总共预测了16个神经肽家族/亚家族，其中14个通过cDNA末端快速扩增被确认为真正的神经肽。肽前体结构和成熟序列的分析表明，amphitrite的某些神经肽是新型同工型，并且具有与昆虫同源的相似特征。预测的神经肽基因的表达谱显示，色素分散激素，SIF酰胺，降钙素和B型尿抑素在塞浦路斯阶段的表达水平最高，而速激肽相关的肽在塞浦路斯和青少年中均被下调。此外，与肽成熟有关的前蛋白转化酶抑制剂有效地延迟了幼虫的变态。实时PCR结果和生物测定的结合表明某些神经肽可能在塞浦路斯定居中发挥重要作用。总体而言，本研究中表征的神经肽/肽激素的新见识应为揭露藤壶幼虫行为和沉降过程的肽能控制提供一个平台。

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