Proteomic Analysis of the Reproductive Organs of the Hermaphroditic Gastropod Lymnaea stagnalis Exposed to Different Endocrine Disrupting Chemicals

Abstract

Many studies have reported perturbations of mollusc reproduction following exposure to low concentrations (ng/L range) of endocrine disrupting chemicals (EDCs). However, the mechanisms of action of these molecules on molluscs are still poorly understood. Investigation of the modifications of protein expression in organisms exposed to chemicals using proteomic methods can provide a broader and more comprehensive understanding of adverse impacts of pollution on organisms than conventional biochemical biomarkers (e.g., heat-shock proteins, metallothioneins, GST, EROD). In this study we have investigated the impacts of four chemicals, which exhibit different endocrine disrupting properties in vertebrates, on the proteome of the hermaphroditic freshwater pulmonate gastropod Lymnaea stagnalis after 21 days of exposure. Testosterone, tributyltin, chlordecone and cyproterone acetate were chosen as tested compounds as they can induce adverse effects on the reproduction of this snail. The 2D-DIGE method was used to identify proteins whose expression was affected by these compounds. In addition to modifying the expression of proteins involved in the structure and function of the cytoskeleton, chemicals had impacts on the expression of proteins involved in the reproduction of L. stagnalis. Exposure to 19.2 µg/L of chlordecone increased the abundance of ovipostatin, a peptide transmitted during mating through seminal fluid, which reduces oviposition in this species. The expression of yolk ferritin, the vitellogenin equivalent in L. stagnalis, was reduced after exposure to 94.2 ng Sn/L of tributyltin. The identification of yolk ferritin and the modification of its expression in snails exposed to chemicals were refined using western blot analysis. Our results showed that the tested compounds influenced the abundance of yolk ferritin in the reproductive organs. Alteration in proteins involved in reproductive pathways (e.g., ovipostatin and yolk ferritin) could constitute relevant evidence of interaction of EDCs with reproductive pathways that are under the control of the endocrine system of L. stagnalis.

Effects of chlordecone on 20-hydroxyecdysone concentration and chitobiase activity in a decapod crustacean, Macrobrachium rosenbergii

“…It has nevertheless been observed that EDCs can also disrupt invertebrate physiology. For example, Giusti et al (2013) have shown that tributyltin affected the production of yolk ferritin (i.e. Vg equivalent) in Lymnaea stagnalis, and Xuereb et al (2011) observed that nonylphenol (estrogenic compound) impacted the expression of a vitellogeninlike gene in the amphipod Gammarus fossarum.…”

Intersex related gene expression profiles in clams Scrobicularia plana: Molecular markers and environmental application

“…COX1 expression has previously been highlighted as differentially regulated, though subunit specific, in clam exhibiting intersex, mussels exposed to metals, and snail exposed to EDCs (Ciocan et al, 2012;Varotto et al, 2013;Giusti et al, 2013). A study into the mechanistic toxicity of imposex in mud snail, Ilyanassa obsoleta, induced using the antifoulant compound irgarol, similarly reports COX1 up regulation (Finnegan et al, 2009).…”

Prednisolone impairs embryonic and posthatching development and shell formation of the freshwater snail, Physa acuta

“…There is a growing body of evidence with respect to the presence of glucocorticoids in the environment adversely affecting aquatic biota . Earlier studies have also showed that environmental contaminants (such as pharmaceutical and personal care products, EDCs, pesticides, and heavy metals) induced malformations or other changes in freshwater snail development and survival . Embryonic developmental toxicity in P. acuta was earlier studied in terms of prominent environmental contaminants such as cadmium , ionic liquids , fluoxetine , and nanoparticles .…”