[en] 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.
Disciplines :
Environmental sciences & ecology
Author, co-author :
Giusti, Arnaud ; Université de Liège - ULiège > Département de Biologie, Ecologie et Evolution > Ecologie animale et écotoxicologie
Leprince, Pierre ; Université de Liège - ULiège > GIGA - Neurosciences
Mazzucchelli, Gabriel ; Université de Liège - ULiège > Center for Analytical Research and Technology (CART)
Lagadic, Laurent
Ducrot, Virginie
Thomé, Jean-Pierre ; Université de Liège - ULiège > Département de Biologie, Ecologie et Evolution > Ecologie animale et écotoxicologie
Joaquim-Justo, Célia ; Université de Liège - ULiège > Département de Biologie, Ecologie et Evolution > Ecologie animale et écotoxicologie
Language :
English
Title :
Proteomic Analysis of the Reproductive Organs of the Hermaphroditic Gastropod Lymnea stagnalis Exposed to Different Endocrine Disrupting Chemicals
Publication date :
November 2013
Journal title :
PLoS ONE
eISSN :
1932-6203
Publisher :
Public Library of Science, San Franscisco, United States - California
Volume :
8
Issue :
11
Pages :
e81086
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
FNRS-FRIA grant FNRS-FRFC grant 2.4.585.10 FRSM grant 3.4559-11
IPCS IPoCS (2002) Global assessment of the state-of-the-science of endocrine disruptors. Geneve, Switzerland. In Damstra T, Barlow S, Bergman A, Kavlock R, Van Der Kraak G
US-EPA (1997) Special report on environmental endocrine disruption: an effects assessment and analyses. report no: EPA/630/R-96/012. p. 120.
Oehlmann J, Schulte-Oehlmann U, Bachmann J, Oetken M, Lutz I et al. (2006) Bisphenol A Induces Superfeminization in the Ramshorn Snail Marisa cornuarietis (Gastropoda: Prosobranchia) at Environmentally Relevant Concentrations. Environ Health Perspect 114: 127-133. PubMed: 16818258.
Oehlmann J, Schulte-Oehlmann U, Tillmann M, Markert B (2000) Effects of Endocrine Disruptors on Prosobranch Snails (Mollusca: Gastropoda) in the Laboratory. Part I: Bisphenol A and Octylphenol as Xeno-Estrogens. Ecotoxicology 9: 383-397. doi:10.1023/A:1008972518019. PubMed: 11214443. (Pubitemid 32108386)
Ortiz-Zarragoitia M, Cajaraville MP (2010) Intersex and oocyte atresia in a mussel population from the Biosphere's Reserve of Urdaibai (Bay of Biscay). Ecotoxicol Environ Saf 73: 693-701. doi:10.1016/j.ecoenv.2010.04.002. PubMed: 20434217.
Gomes T, Gonzalez-Rey M, Bebianno MJ (2009) Incidence of intersex in male clams Scrobicularia plana in the Guadiana Estuary (Portugal). Ecotoxicology 18: 1104-1109. doi:10.1007/s10646-009-0359-5. PubMed: 19633956.
Bauer B, Fioroni P, Schulte-Oehlmann U, Oehlmann J, Kalbfus W (1997) The use of Littorina littorea for tributyltin (TBT) effect monitoring - Results from the German TBT survey 1994/1995 and laboratory experiments. Environ Pollut 96: 299-309. doi:10.1016/S0269-7491(97)00049-3. PubMed: 15093396. (Pubitemid 27390440)
Oehlmann J, Bauer B, Minchin D, Schulte-Oehlmann U, Fioroni P et al. (1998) Imposex in Nucella lapillus and intersex in Littorina littorea: interspecific comparison of two TBT-induced effects and their geographical uniformity. Hydrobiologia 378: 199-213. doi:10.1023/A:1003218411850.
Duft M, Schulte-Oehlmann U, Weltje L, Tillmann M, Oehlmann J (2003) Stimulated embryo production as a parameter of estrogenic exposure via sediments in the freshwater mudsnail Potamopyrgus antipodarum. Aquat Toxicol 64: 437-449. doi:10.1016/S0166-445X(03)00102-4. PubMed: 12878414.
Tillmann M, Schulte-Oehlmann U, Duft M, Markert B, Oehlmann J (2001) Effects of Endocrine Disruptors on Prosobranch Snails (Mollusca: Gastropoda) in the Laboratory. Part III: Cyproterone Acetate and Vinclozolin as Antiandrogens. Ecotoxicology 10: 373-388. doi: 10.1023/A:1012279231373. PubMed: 11759570. (Pubitemid 33070882)
Czech P, Weber K, Dietrich DR (2001) Effects of endocrine modulating substances on reproduction in the hermaphroditic snail Lymnaea stagnalis L. Aquat Toxicol 53: 103-114. doi:10.1016/S0166-445X(00)00169-7. PubMed: 11311387.
Ducrot V, Teixeira-Alves M, Lopes C, Delignette-Muller M-L, Charles S et al. (2010) Development of partial life-cycle experiments to assess the effects of endocrine disruptors on the freshwater gastropod Lymnaea stagnalis: a case-study with vinclozolin. Ecotoxicology 19: 1312-1321. doi:10.1007/s10646- 010-0518-8. PubMed: 20623335.
Leung KMY, Grist EPM, Morley NJ, Morritt D, Crane M (2007) Chronic toxicity of tributyltin to development and reproduction of the European freshwater snail Lymnaea stagnalis (L.). Chemosphere 66: 1358-1366. (Pubitemid 44854082)
Giusti A, Barsi A, Dugué M, Collinet M, Thomé J-P et al. (2013) Reproductive impacts of tributyltin (TBT) and triphenyltin (TPT) in the hermaphroditic freshwater gastropod Lymnaea stagnalis. Environ Toxicol Chem 32: 1552-1560. PubMed: 23450754.
Geraerts WPM (1976) The control of ovulation in the hermaphroditic freshwater snail Lymnaea stagnalis by the neurohormone of the caudodorsal cells. Gen Comp Endocrinol 28: 350-357. doi:10.1016/0016-6480(76)90187-8. PubMed: 947220.
Koene J (2010) Neuro-endocrine control of reproduction in hermaphroditic freshwater snails: mechanisms and evolution. Front. Behav Neurosci 4: 167-167.
Rittschof D, McClellan-Green P (2005) Molluscs as multidisciplinary models in environment toxicology. Mar Pollut Bull 50: 369-373. doi: 10.1016/j.marpolbul.2005.02.008. PubMed: 15823298.
Bouétard A, Noirot C, Besnard A-L, Bouchez O, Choisne D et al. (2012) Pyrosequencing-based transcriptomic resources in the pond snail Lymnaea stagnalis, with a focus on genes involved in molecular response to diquat-induced stress. Ecotoxicology 21: 2222-2234. doi:10.1007/s10646-012-0977- 1. PubMed: 22814884.
OECD (2010) Detailed Review Paper on Mollusc Life-Cycle Toxicity Testing. Paris, France: Organisation for Economic Co-operation and Development Environment Directorate. 182 p
Cao A, Fuentes J, Comesana P, Casas SM, Villalba A (2009) A proteomic approach envisaged to analyse the bases of oyster tolerance/resistance to bonamiosis. Aquaculture 295: 149-156. doi:10.1016/j.aquaculture.2009.06.044.
Wright PC, Noirel J, Ow SY, Fazeli A (2012) A review of current proteomics technologies with a survey on their widespread use in reproductive biology investigations. Theriogenology 77: 738-765.e752
Sanchez BC, Ralston-Hooper K, Sepúlveda MS (2011) Review of recent proteomic applications in aquatic toxicology. Environ Toxicol Chem 30: 274-282. doi:10.1002/etc.402. PubMed: 21072841.
Rodríguez-Ortega MJ, Grøsvik BE, Rodríguez-Ariza A, Goksøyr A, López-Barea J (2003) Changes in protein expression profiles in bivalve molluscs (Chamaelea gallina) exposed to four model environmental pollutants. Proteomics 3: 1535-1543. doi:10.1002/pmic.200300491. PubMed: 12923779.
Apraiz I, Mi J, Cristobal S (2006) Identification of Proteomic Signatures of Exposure to Marine Pollutants in Mussels (Mytilus edulis). Mol Cell Proteomics 5: 1274-1285. doi:10.1074/mcp.M500333-MCP200. PubMed: 16603574. (Pubitemid 44103398)
Benninghoff AD (2007) Toxicoproteomics - The Next Step in the Evolution of Environmental Biomarkers? Toxicol Sci 95: 1-4. PubMed: 17209232.
Dowling VA, Sheehan D (2006) Proteomics as a route to identification of toxicity targets in environmental toxicology. Proteomics 6: 5597-5604. doi:10.1002/pmic.200600274. PubMed: 16972288. (Pubitemid 44683223)
Campos A, Tedesco S, Vasconcelos V, Cristobal S (2012) Proteomic research in bivalves: Towards the identification of molecular markers of aquatic pollution. J Proteomics 75: 4346-4359. doi:10.1016/j.jprot.2012.04.027. PubMed: 22579653.
Ankley GT, Bencic DC, Breen MS, Collette TW, Conolly RB et al. (2009) Endocrine disrupting chemicals in fish: Developing exposure indicators and predictive models of effects based on mechanism of action. Aquat Toxicol 92: 168-178. doi:10.1016/j.aquatox.2009.01.013. PubMed: 19261338.
Shepard JL, Olsson B, Tedengren M, Bradley BP (2000) Protein expression signatures identified in Mytilus edulis exposed to PCBs, copper and salinity stress. Mar Environ Res 50: 337-340. doi:10.1016/S0141-1136(00)00065-9. PubMed: 11460714.
Thompson EL, Taylor DA, Nair SV, Birch G, Haynes PA et al. (2012) Proteomic discovery of biomarkers of metal contamination in Sydney Rock oysters (Saccostrea glomerata). Aquat Toxicol 109: 202-212. doi:10.1016/j.aquatox.2011. 09.018. PubMed: 22030410.
Zhou J, Cai Z-H, Li L, Gao Y-F, Hutchinson TH (2010) A proteomics based approach to assessing the toxicity of bisphenol A and diallyl phthalate to the abalone (Haliotis diversicolor supertexta). Chemosphere 79: 595-604. doi:10.1016/j.chemosphere.2010.01.052. PubMed: 20189630.
Giusti A, Ducrot V, Joaquim-Justo C, Lagadic L (2013) Testosterone levels and fecundity in the hermaphroditic aquatic snail Lymnaea stagnalis exposed to testosterone and endocrine disruptors. Environ Toxicol Chem 32: 1740-1745. doi:10.1002/etc.2234. PubMed: 23564527.
Lavado R, Janer G, Porte C (2006) Steroid levels and steroid metabolism in the Mussel Mytilus edulis: The modulating effect of dispersed crude oil and alkylphenols. Aquat Toxicol 78: S65-S72. doi: 10.1016/j.aquatox.2006.02.018. PubMed: 16600398.
Gooding MP, LeBlanc GA (2004) Seasonal variation in the regulation of testosterone levels in the eastern mud snail (Ilyanassa obsoleta). Invertebr Biol 123: 237-243. (Pubitemid 39474460)
Gauthier-Clerc S, Pellerin J, Amiard JC (2006) Estradiol-17β and testosterone concentrations in male and female Mya Arenaria (Mollusca bivalvia) during the reproductive cycle. Gen Comp Endocrinol 145: 133-139. doi:10.1016/j.ygcen.2005.08.004. PubMed: 16197945. (Pubitemid 43005047)
Liu WK, Li Q, Kong L (2008) Estradiol-17β and testosterone levels in the cockle Fulvia mutica during the annual reproductive cycle. New Zeal J Mar Fresh 42: 417-424. doi:10.1080/00288330809509970.
Wang C, Croll RP (2004) Effects of sex steroids on gonadal development and gender determination in the sea scallop, Placopecten magellanicus. Aquaculture 238: 483-498. doi:10.1016/j.aquaculture.2004.05.024. (Pubitemid 39123663)
Wang C, Croll RP (2006) Effects of sex steroids on spawning in the sea scallop, Placopecten magellanicus. Aquaculture 256: 423-432. doi:10.1016/j.aquaculture.2006.01.017. (Pubitemid 43767180)
Wang C, Croll RP (2003) Effects of sex steroids on in vitro gamete release in the sea scallop, Placopecten magellanicus. Invert Reprod Dev 44: 89-100. doi:10.1080/07924259.2003.9652559. (Pubitemid 37546142)
Fang H, Tong W, Branham WS, Moland CL, Dial SL et al. (2003) Study of 202 Natural, Synthetic, and Environmental Chemicals for Binding to the Androgen Receptor. Chem Res Toxicol 16: 1338-1358. doi:10.1021/tx030011g. PubMed: 14565775. (Pubitemid 37315223)
Hammond B, Katzenellenbogen BS, Krauthammer N, McConnell J (1979) Estrogenic activity of the insecticide chlordecone (Kepone) and interaction with uterine estrogen receptors. Proc Natl Acad Sci USA 76: 6641-6645. doi:10.1073/pnas.76.12.6641. PubMed: 93289.
Donohoe RM, Curtis LR (1996) Estrogenic activity of chlordecone, o,p′-DDT and o,p′-DDE in juvenile rainbow trout: induction of vitellogenesis and interaction with hepatic estrogen binding sites. Aquat Toxicol 36: 31-52. doi:10.1016/S0166-445X(96)00799-0.
Horiguchi T, Lee J-H, Park J-C, Cho H-S, Shiraishi H et al. (2012) Specific accumulation of organotin compounds in tissues of the rock shell, Thais clavigera. Mar Environ Res 76: 56-62. doi:10.1016/j.marenvres.2011.09.002. PubMed: 21955599.
Sternberg RM, Gooding MP, Hotchkiss AK, LeBlanc GA (2010) Environmental-endocrine control of reproductive maturation in gastropods: Implications for the mechanism of tributyltin-induced imposex in prosobranchs. Ecotoxicology 19: 4-23. doi:10.1007/s10646-009-0397-z. PubMed: 19653098.
Benachour N, Moslemi S, Sipahutar H, Seralini G-E (2007) Cytotoxic effects and aromatase inhibition by xenobiotic endocrine disrupters alone and in combination. Toxicol Appl Pharmacol 222: 129-140. doi:10.1016/j.taap.2007.03. 033. PubMed: 17599374. (Pubitemid 47103185)
Saitoh M, Yanase T, Morinaga H, Tanabe M, Mu Y-M et al. (2001) Tributyltin or Triphenyltin Inhibits Aromatase Activity in the Human Granulosa-like Tumor Cell Line KGN. Biochem Bioph Res Co. 289: 198-204 PubMed: 11708799. (Pubitemid 33108814)
Raudrant D, Rabe T (2003) Progestogens with antiandrogenic properties. Drugs 63: 463-492. doi:10.2165/00003495-200363050-00003. PubMed: 12600226. (Pubitemid 36337727)
Coutellec M-A, Lagadic L (2006) Effects of Self-Fertilization, Environmental Stress and Exposure to Xenobiotics on Fitness-Related Traits of the Freshwater Snail Lymnaea stagnalis. Ecotoxicology 15: 199-213. doi:10.1007/s10646-005-0049-x. PubMed: 16425104.
OECD (2000) Guidance document on aquatic toxicity testing of difficult substances and mixtures. Paris, France: Organisation for Economic Co-operation and Development Environment Directorate. 53 p
ISO ISO (2004) ISO TC 147/SC2 Water quality - Determination of selected organotin compounds - Gas chromatographic method. Geneva, Switzerland: ISO. p. 31.
Belgers JDM, Aalderink GH, Arts GHP, Brock TCM (2011) Can time-weighted average concentrations be used to assess the risks of metsulfuron-methyl to Myriophyllum spicatum under different time-variable exposure regimes? Chemosphere 85: 1017-1025. doi:10.1016/j.chemosphere.2011.07.025. PubMed: 21875741.
Leroy D, Haubruge E, De Pauw E, Thomé JP, Francis F (2010) Development of ecotoxicoproteomics on the freshwater amphipod Gammarus pulex: Identification of PCB biomarkers in glycolysis and glutamate pathways. Ecotoxicol Environ Saf 73: 343-352. doi:10.1016/j.ecoenv.2009.11.006. PubMed: 20015547.
Francis F, Guillonneau F, Leprince P, De Pauw E, Haubruge E et al. (2010) Tritrophic interactions among Macrosiphum euphorbiae aphids, their host plants and endosymbionts: Investigation by a proteomic approach. J Insect Physiol 56: 575-585. doi:10.1016/j.jinsphys.2009.12.001. PubMed: 19962988.
Leung PTY, Wang Y, Mak SST, Ng WC, Leung KMY (2011) Differential proteomic responses in hepatopancreas and adductor muscles of the green-lipped mussel Perna viridis to stresses induced by cadmium and hydrogen peroxide. Aquat Toxicol 105: 49-61. doi:10.1016/j.aquatox.2011.05.010. PubMed: 21684241.
Berth M, Moser FM, Kolbe M, Bernhardt J (2007) The state of the art in the analysis of two-dimensional gel electrophoresis images. Appl Microbiol Biotechnol 76: 1223-1243. doi:10.1007/s00253-007-1128-0. PubMed: 17713763. (Pubitemid 47459790)
Collodoro M, Lemaire P, Eppe G, Bertrand V, Dobson R et al. (2012) Identification and quantification of concentration-dependent biomarkers in MCF-7/BOS cells exposed to 17β-estradiol by 2-D DIGE and label-free proteomics. J Proteomics 75: 4555-4569. doi:10.1016/j.jprot.2012.04.032. PubMed: 22580036.
Aldridge GM, Podrebarac DM, Greenough WT, Weiler IJ (2008) The use of total protein stains as loading controls: An alternative to high-abundance single-protein controls in semi-quantitative immunoblotting. J Neurosci Methods 172: 250-254. doi:10.1016/j.jneumeth.2008.05.003. PubMed: 18571732.
Romero-Calvo I, Ocón B, Martínez-Moya P, Suárez MD, Zarzuelo A et al. (2010) Reversible Ponceau staining as a loading control alternative to actin in Western blots. Anal Biochem 401: 318-320. doi:10.1016/j.ab.2010.02.036. PubMed: 20206115.
D'Agostino RB, Pearson ES (1973) Tests for departure from normality. Empirical results for the distributions of b2 and √b1. Biometrika 60: 613-622. doi:10.1093/biomet/60.3.613.
Bartlett MS, Kendall DG (1946) The Statistical Analysis of Variance-Heterogeneity and the Logarithmic Transformation. Suppl J Royal Stat Soc 8: 128-138.
Cox DN, Chao A, Lin H (2000) piwi encodes a nucleoplasmic factor whose activity modulates the number and division rate of germline stem cells. Development 127: 503-514. PubMed: 10631171. (Pubitemid 30093398)
Koene JM, Sloot W, Montagne-Wajer K, Cummins SF, Degnan BM et al. (2010) Male Accessory Gland Protein Reduces Egg Laying in a Simultaneous Hermaphrodite. PLOS ONE 5: e10117. doi:10.1371/journal.pone.0010117. PubMed: 20404934.
Bottke W, Burschyk M, Volmer J (1988) On the origin of the yolk protein ferritin in snails. Dev Genes Evol 197: 377-382. (Pubitemid 19010656)
Bottke W, Sinha I (1979) Ferritin as an exogenous yolk protein in snails. Dev Genes Evol 186: 71-75. (Pubitemid 9115415)
Bradley BP, Shrader EA, Kimmel DG, Meiller JC (2002) Protein expression signatures: an application of proteomics. Mar Environ Res 54: 373-377. doi:10.1016/S0141-1136(02)00115-0. PubMed: 12408590. (Pubitemid 35286451)
Dondero F, Negri A, Boatti L, Marsano F, Mignone F et al. (2010) Transcriptomic and proteomic effects of a neonicotinoid insecticide mixture in the marine mussel (Mytilus galloprovincialis, Lam.). Sci Total Eviron 408: 3775-3786.
Thompson EL, Taylor DA, Nair SV, Birch G, Hose GC et al. (2012) Proteomic analysis of Sydney Rock oysters (Saccostrea glomerata) exposed to metal contamination in the field. Environ Pollut 170: 102-112. doi:10.1016/j.envpol. 2012.06.015. PubMed: 22771357.
Chora S, Starita-Geribaldi M, Guigonis J-M, Samson M, Roméo M et al. (2009) Effect of cadmium in the clam Ruditapes decussatus assessed by proteomic analysis. Aquat Toxicol 94: 300-308. doi:10.1016/j.aquatox.2009.07. 014. PubMed: 19716188.
Miura Y, Kano M, Abe K, Urano S, Suzuki S et al. (2005) Age-dependent variations of cell response to oxidative stress: Proteomic approach to protein expression and phosphorylation. Electrophoresis 26: 2786-2796. doi:10.1002/elps.200500172. PubMed: 15966013. (Pubitemid 41078988)
McDonagh B, Tyther R, Sheehan D (2005) Carbonylation and glutathionylation of proteins in the blue mussel Mytilus edulis detected by proteomic analysis and Western blotting: Actin as a target for oxidative stress. Aquat Toxicol 73: 315-326. doi:10.1016/j.aquatox.2005.03.020. PubMed: 15869813. (Pubitemid 40797584)
Pascoal S, Carvalho G, Vasieva O, Hughes R, Cossins AR et al. (2012) Transcriptomics and in vivo tests reveal novel mechanisms underlying endocrine disruption in an ecological sentinel, Nucella lapillus. Mol Ecol.
Manduzio H, Cosette P, Gricourt L, Jouenne T, Lenz C et al. (2005) Proteome modifications of blue mussel (Mytilus edulis L.) gills as an effect of water pollution. Proteomics 5: 4958-4963. doi:10.1002/pmic.200401328. PubMed: 16267813. (Pubitemid 43016914)
Apraiz I, Cajaraville MP, Cristobal S (2009) Peroxisomal proteomics: Biomonitoring in mussels after the Prestige's oil spill. Mar Pollut Bull 58: 1815-1826. doi:10.1016/j.marpolbul.2009.08.004. PubMed: 19732914.
Zhou J, Cai Z-H, Zhu X-S, Li L, Gao Y-F (2010) Innate immune parameters and haemolymph protein expression profile to evaluate the immunotoxicity of tributyltin on abalone (Haliotis diversicolor supertexta). Dev Comp Immunol 34: 1059-1067. doi:10.1016/j.dci.2010.05.006. PubMed: 20488199.
Cox DN, Chao A, Baker J, Chang L, Qiao D et al. (1998) A novel class of evolutionarily conserved genes defined by piwi are essential for stem cell self-renewal. Gen Dev 12: 3715-3727. doi:10.1101/gad.12.23.3715. (Pubitemid 29019652)
Zhou X, Liao Z, Jia Q, Cheng L, Li F (2007) Identification and characterization of Piwi subfamily in insects. Biochem Bioph Res Co 362: 126-131 (Pubitemid 47296302)
Beyret E, Lin H (2011) Pinpointing the expression of piRNAs and function of the PIWI protein subfamily during spermatogenesis in the mouse. Dev Biol 355: 215-226. doi:10.1016/j.ydbio.2011.04.021. PubMed: 21539824.
Wang G, Reinke V (2008) A C. elegans Piwi, PRG-1, Regulates 21U-RNAs during Spermatogenesis. Curr Biol 18: 861-867. doi:10.1016/j.cub.2008.05.009. PubMed: 18501605.
Hoffer JNA (2012) Quantifying Sexual Conflict in a Simultaneous Hermaphrodite. Amsterdam: Vrije Universiteit.
Bottke W (1986) Immuno-localization of ferritin polypeptides in oocytes and somatic tissue of the freshwater snails Lymnaea stagnalis L. and Planorbarius corneus L. Cell Tissue Res 243: 397-404.
Agnese M, Verderame M, De Meo E, Prisco M, Rosati L et al. (2013) A network system for vitellogenin synthesis in the mussel Mytilus galloprovincialis (L.). J Cell Physiol 228: 547-555.
Björkblom C, Mustamäki N, Olsson P-E, Katsiadaki I, Wiklund T (2011) Assessment of reproductive biomarkers in three-spined stickleback (Gasterosteus aculeatus) from sewage effluent recipients. Environ Toxicol 28: 229-237. PubMed: 21656640.
Purdom CE, Hardiman PA, Bye VVJ, Eno NC, Tyler CR et al. (1994) Estrogenic Effects of Effluents from Sewage Treatment Works. Chem Ecol 8: 275-285. doi:10.1080/02757549408038554.
Harries JE, Janbakhsh A, Jobling S, Matthiessen P, Sumpter JP et al. (1999) Estrogenic potency of effluent from two sewage treatment works in the United Kingdom. Environ Toxicol Chem 18: 932-937. doi:10.1002/etc.5620180517. (Pubitemid 29186246)
Sumpter JP, Jobling S (1995) Vitellogenesis as a biomarker for estrogenic contamination of the aquatic environment. Environ Health Perspect 103 Suppl 7: 173-178. doi:10.2307/3432529. PubMed: 8593867.
OECD (2009) Test No. 229: Fish Short Term Reproduction Assay. OECD Publishing.
Andrew MN, Dunstan RH, O'Connor WA, Van Zwieten L, Nixon B et al. (2008) Effects of 4-nonylphenol and 17α-ethynylestradiol exposure in the Sydney rock oyster, Saccostrea glomerata: Vitellogenin induction and gonadal development. Aquat Toxicol 88: 39-47. doi:10.1016/j.aquatox.2008.03.003. PubMed: 18453011.
Andrew MN, O'Connor WA, Dunstan RH, MacFarlane GR (2010) Exposure to 17α-ethynylestradiol causes dose and temporally dependent changes in intersex, females and vitellogenin production in the Sydney rock oyster. Ecotoxicology 19: 1440-1451. doi:10.1007/s10646-010-0529-5. PubMed: 20700763.
Matozzo V, Marin MG (2008) Can 17-estradiol induce vitellogenin-like proteins in the clam Tapes philippinarum? Environ Toxicol Chem 26: 38-44.
Andrew-Priestley MN, O'Connor WA, Dunstan RH, Van Zwieten L, Tyler T et al. (2012) Estrogen mediated effects in the Sydney rock oyster, Saccostrea glomerata, following field exposures to sewage effluent containing estrogenic compounds and activity. Aquat Toxicol 120-121: 99-108. PubMed: 22673404.
Zhang H, Pan L, Zhang L (2012) Molecular cloning and characterization of estrogen receptor gene in the Scallop Chlamys farreri: Expression profiles in response to endocrine disrupting chemicals. Comp Biochem Physiol C Toxicol Pharmacol 156: 51-57. doi:10.1016/j.cbpc.2012.03.007. PubMed: 22507668.
Matozzo V, Gagné F, Marin MG, Ricciardi F, Blaise C (2008) Vitellogenin as a biomarker of exposure to estrogenic compounds in aquatic invertebrates: A review. Environ Int 34: 531-545. doi:10.1016/j.envint.2007.09. 008. PubMed: 18029015. (Pubitemid 351681916)
Damodaran S, Wood TD, Nagarajan P, Rabin RA (2007) Evaluating Peptide Mass Fingerprinting-based Protein Identification. Genomics, Proteomics Bioinformatics 5: 152-157. doi:10.1016/S1672-0229(08)60002-9. PubMed: 18267296. (Pubitemid 351215114)
OECD (2012) Guidance document on standardised test guidelines for evaluating chemicals for endocrine disruption. Paris: Organisation for Economic Co-operation and Development.