Abstract

Because of their crucial role in ecotoxicological risk assessment, amphipods (Crustacea) are commonly employed as model species in a wide range of studies. However, despite their ecological importance, their genome has not yet been completely annotated and molecular mechanisms underlying key pathways, such as the serotonin pathway, in development of ecotoxicological biomarkers of exposure to neuroactive pharmaceuticals are still poorly understood. Furthermore, genetic similarities and discrepancies with other model arthropods (e.g., Drosophila melanogaster) have not been completely clarified. In this report, we present a new transcriptome assembly of Gammarus fossarum, an important amphipod species, widespread in Central Europe. RNA-Seq with Illumina HiSeq technology was used to analyse samples extracted from total internal tissues. We used the Trinity and Trinotate software suites for transcriptome assembly and annotation, respectively. The quality of this assembly and the affiliated targeted homology searches greatly enrich the molecular knowledge on this species. Because of the lack of publicly available molecular information on the serotonin pathway, we also highlighted sequence homologies and divergences of the genes encoding the serotonin pathway components of the well-annotated arthropod D. melanogaster, and Crustacea with the corresponding genes of our assembly. An inferior number of hits was found when running a BLAST analysis of both D. melanogaster and Crustacea mRNA sequences encoding serotonin receptors available in GenBank against the total assembly, compared to other serotonin pathway components. A lack of information on important components for serotonin biosynthesis and vesicle endocytosis (i.e., tryptophan hydroxylase and vesicular monoamine transporter) in Crustacea was also brought to light. Our results will provide an extensive transcriptional resource for this important species in ecotoxicological risk assessment and highlight the need for a more detailed categorization of neuronal pathways components in invertebrates.

Highlights

  • Identifying new sentinel species and acquiring molecular data for biomarker analyses are fundamental objectives in today’s ecotoxicological research

  • Sequencing generated a total of 325,393,762 paired end reads across the 20 samples

  • Considering that the historical contribution of Crustacea in synaptic physiology is unsurpassed (Atwood, 1976; Wiese, 2013) and much is still unclear concerning the synaptic molecular mechanisms within this taxon (Wu and Cooper, 2012), we carried out a manual annotation of the genes encoding the serotonin-related molecular components present in the final assembly. We focused on this particular neurotransmitter because of the lack of general molecular information in invertebrates in contrast with the wide range of documented behavioural and transcriptional effects of antidepressants released in aquatic environments (Nentwig, 2007; Guler and Ford, 2010; Bossus et al, 2014; Ford and Fong, 2016; Estévez-Calvar et al, 2017)

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Summary

Introduction

Identifying new sentinel species and acquiring molecular data for biomarker analyses are fundamental objectives in today’s ecotoxicological research. In research on aquatic species for instance, -omics platforms have allowed the development of new biomarkers of exposure to a considerably higher number of chemicals, allowing an even more precise description of the molecular effects in response to the exposure to a range of ecologically dangerous substances (Hauser-Davis et al, 2012; Jayapal, 2012; Barrera and Ariza, 2017; Poynton et al, 2018). With particular regard to freshwater species, numerous studies have been carried out to evaluate the molecular effects of a large spectrum of substances unremoved by wastewater treatment plants (WWTPs).

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