Identification of the first neuropeptides from the enigmatic hexapod order Protura

The Hexapoda consists of two classes, the Entognatha and the Insecta, with the former group considered basal to the latter. The Protura is a basal order within the Entognatha, the members of which are minute soil dwellers first identified in the early 20th century. Recently, a transcriptome shotgun assembly (TSA) was generated for the proturan Acerentomon sp., providing the first significant molecular resource for this enigmatic hexapod order. As part of an ongoing effort to predict peptidomes for little studied members of the Arthropoda, we have mined this TSA dataset for transcripts encoding putative neuropeptide precursors and predicted the structures of mature peptides from the deduced proteins. Forty-seven peptide-encoding transcripts were mined from the Acerentomon TSA dataset, with 202 distinct peptides predicted from them. The peptides identified included isoforms of adipokinetic hormone, adipokinetic hormone-corazonin-like peptide, allatostatin A, allatostatin B, allatostatin C, allatotropin, bursicon α, bursicon β, CCHamide, corazonin, crustacean cardioactive peptide, crustacean hyperglycemic hormone/ion transport peptide, diuretic hormone 31, diuretic hormone 44, ecdysis-triggering hormone, eclosion hormone, FMRFamide-like peptide, GSEFLamide, insulin-like peptide, intocin, leucokinin, myosuppressin, neuropeptide F, orcokinin, proctolin, pyrokinin, RYamide, short neuropeptide F, SIFamide, sulfakinin and tachykinin-related peptide; these are the first neuropeptides described from any proturan. Comparison of the Acerentomon precursors and mature peptides with those from other arthropods revealed features characteristic of both the insects and the crustaceans, which is consistent with the hypothesized phylogenetic position of the Protura within the Pancrustacea, i.e. at or near the point of divergence of the hexapods from the crustaceans.