Vitellogenin-like Proteins among Invertebrate Species Diversity: Potential of Proteomic Mass Spectrometry for Biomarker Development

Abstract

Cost-effective methodologies along with cross-species applicability constitute key points for biomarker development in ecotoxicology. With the advent of cheaper affordable genomic techniques and high throughput sequencing, omics tools could facilitate the assessment of effects of environmental contaminants for all taxa biodiversity. We assessed the potential of absolute quantification of proteins using mass spectrometry to develop vitellogenin (Vg)-like protein assays for invertebrates. We used available sequences in public databases to rapidly identify Vg-proteotypic peptides in seven species from different main taxa of protostome invertebrates (mollusk bivalves, crustacean amphipods, branchiopods, copepods and isopods, and insect diptera). Functional validation was performed by comparing proteomic signals from reproductive female tissue samples and negative controls (male or juvenile tissues). In a second part, we demonstrate in gammarids, daphnids, drosophilids, and gastropods that the assay validated in Vg-sequenced species can be applied to Vg-unsequenced species thanks to the evolutionary conservation of Vg-proteotypic peptide motifs. Finally, we discuss the relevance of mass spectrometry for biomarker development (specific measurement, rapid development, transferability across species). Our study supplies an illustration of the promising strategy to address the challenge of biodiversity in ecotoxicology, which consists in employing omics tools from comparative and evolutionary perspectives.

Application of a redox‐proteomics toolbox to Daphnia magna challenged with model pro‐oxidants copper and paraquat

“…The mechanistic significance of changes in vtg expression or maturation is not known in crustaceans, although it is of interest that the changes in protein content and carbonylation levels observed in the present study vary markedly from those of a previous study on silver nanoparticles and silver nitrate. This is in agreement with previous publications indicating that the vtg subproteome is an easily measured marker of physiological change in vertebrates and invertebrates alike, yielding specific responses to stressors . This may facilitate identification of protein expression signatures (PES) with potential to differentiate stressors.…”

Proteomic evaluation of citrate-coated silver nanoparticles toxicity in Daphnia magna

“…29 Yet work has been completed to establish vitellogenins as an easily applicable biomarker between invertebrate species, including daphnids. 30 In correspondence to this, vitellogenin has been proposed as a marker of general stress in the zebra sh, especially when egg or embryo development is followed …”

Vitellogenin is not an appropriate biomarker of feminisation in a Crustacean

“…The primary site of Vtg transcription appears to be the ovary, with several studies having highlighted extraovarian synthesis of Vtg in the hepatopancreas of some species (Fainzilber et al, 1992;Tsutsui et al, 2000;Mak et al, 2005;Raviv et al, 2006;Tsang et al, 2003). Scientists have also developed and utilised Vtg as a biomarker to determine whether feminisation and/or reproductive endocrine disruption is occurring in invertebrates (Matozzo et al, 2008;Simon et al, 2010;Jubeaux et al, 2012a;Jubeaux et al, 2012b). Such studies have included a diverse range of crustaceans, such as Daphnia, mysids, amphipods, crabs, crayfish, lobsters and various shrimps and prawns (Fainzilber et al, 1992;Lee and Noone, 1994;Sagi et al, 1999;Tsutsui et al, 2000;Allen et al, 2002;Tsang et al, 2003;Ghekiere et al, 2004;Volz and Chandler, 2004;Ghekiere et al, 2005;Mak et al, 2005;Sanders et al, 2005;Zapata-Perez et al, 2005;Ghekiere et al, 2006;Raviv et al, 2006;Simon et al, 2010;Hannas et al, 2011;Xuereb et al, 2011;García, and Heras, 2012;Jubeaux et al, 2012b;Jubeaux et al, 2012c).…”

“…A BLAST analysis (tblastx) against nonredundant sequences in GenBank reveals the sequence is most similar to multiple unannotated 'transcribed RNA sequences' (eg GAKD01006973.1 and GAJQ01003147.1)belonging to freshwater amphipods Melita plumulosa and Hyalella azteca. The annotated sequence with highest level of similarity (e-value 2e-20) to Vtg1 is a partial coding sequence described as 'vitellogenin-like protein precursor' (GU985184.1) isolated from the freshwater amphipod Gammarus fossarum, a sequence that has been characterised for its potential application in studies of endocrine disruption(Simon et al, 2010;Xuereb et al, 2011;Jubeaux et al, 2012a;Jubeaux et al, 2012b). The Vtg2 sequence consists of a 643 bp open reading frame predicted to encode a 214 amino acid sequence.…”