In silico prediction of the G-protein coupled receptors expressed during the metamorphic molt of Sagmariasus verreauxi (Crustacea: Decapoda) by mining transcriptomic data: RNA-seq to repertoire

• Published in: General and comparative endocrinology Vol. 228; pp. 111 - 127
• Main Authors: Buckley, Sean J., Fitzgibbon, Quinn P., Smith, Gregory G., Ventura, Tomer
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.03.2016
• Subjects: Animals; Arthropoda; Computer Simulation; Crustacean hyperglycemic hormone (CHH); Endocrinology & Metabolism; G-protein coupled receptor (GPCR); Gene Expression Profiling; Gene Expression Regulation; High-Throughput Nucleotide Sequencing - methods; Metamorphosis; Metamorphosis, Biological - genetics; Molting - genetics; Palinuridae - classification; Palinuridae - genetics; Phylogeny; Pontophilus spinosus; Real-Time Polymerase Chain Reaction; Receptors, G-Protein-Coupled - genetics; RNA - genetics; Sagmariasus verreauxi; Transcriptomic expression; Transcriptomic expression; G-protein coupled receptor (GPCR); Sagmariasus verreauxi; Metamorphosis; Crustacean hyperglycemic hormone (CHH)
• ISSN: 0016-6480; 1095-6840
• DOI: 10.1016/j.ygcen.2016.02.001

•Digital repertoire of GPCRs expressed in metamorphic Sagmariasus verreauxi.•Neuropeptide GPCRs lack obvious metamorphic differential expression.•A biogenic amine-related receptor expressed predominantly in the testes.•Modelling suggests conserved CHH-like receptor binding pocket. Against a backdrop of food insecurity, the farming of decapod crustaceans is a rapidly expanding and globally significant source of food protein. Sagmariasus verreauxi spiny lobster, the subject of this study, are decapods of underdeveloped aquaculture potential. Crustacean neuropeptide G-protein coupled receptors (GPCRs) mediate endocrine pathways that are integral to animal fecundity, growth and survival. The potential use of novel biotechnologies to enhance GPCR-mediated physiology may assist in improving the health and productivity of farmed decapod populations. This study catalogues the GPCRs expressed in the early developmental stages, as well as adult tissues, with a view to illuminating key neuropeptide receptors. De novo assembled contiguous sequences generated from transcriptomic reads of metamorphic and post metamorphic S. verreauxi were filtered for seven transmembrane domains, and used as a reference for iterative re-mapping. Subsequent putative GPCR open reading frames (ORFs) were BLAST annotated, categorised, and compared to published orthologues based on phylogenetic analysis. A total of 85 GPCRs were digitally predicted, that represented each of the four arthropod subfamilies. They generally displayed low-level and non-differential metamorphic expression with few exceptions that we examined using RT-PCR and qPCR. Two putative CHH-like neuropeptide receptors were annotated. Three dimensional structural modelling suggests that these receptors exhibit a conserved extracellular ligand binding pocket, providing support to the notion that these receptors co-evolved with their ligands across Decapoda. This perhaps narrows the search for means to increase productivity of farmed decapod populations.