{"refrec":{"BRefID":225700,"RR":"De Wilde, R.; Swevers, L.; Soin, T.; Christiaens, O.; Roug\u00e9, P.; Cooreman, K.; Janssen, C.R.; Smagghe, G.<\/b> (2013). Cloning and functional analysis of the ecdysteroid receptor complex in the opossum shrimp Neomysis integer<\/i> (Leach, 1814). Aquat. Toxicol. 130-131<\/i>: 31-40. http:\/\/dx.doi.org\/10.1016\/j.aquatox.2012.12.011<\/a>","BEntID":217424,"PublicFlag":1,"CheckedFlag":1,"wosflag":1,"vabbflag":null,"RefStringPartII":". Aquat. Toxicol. 130-131<\/i>: 31-40. http:\/\/dx.doi.org\/10.1016\/j.aquatox.2012.12.011<\/a>","DocTypID":8,"DocType":"Journal article","MarineFlag":1,"FreshFlag":0,"BrackishFlag":0,"TerrestrialFlag":0,"Authorstring":"De Wilde, R.; Swevers, L.; Soin, T.; Christiaens, O.; Roug\u00e9, P.; Cooreman, K.; Janssen, C.R.; Smagghe, G.","OrigTitleTranslFlag":0,"Authorstringtrunc":"De Wilde, R. et al.<\/i>","Englishabstract":"In this paper, the non-target effects of tebufenozide were evaluated on the estuarine crustacean, the opposum shrimp Neomysis integer<\/i> (Leach, 1814). Tebufenozide is a synthetic non-steroidal ecdysone agonist insecticide and regarded as potential endocrine-disrupting chemical (EDC). N. integer<\/i> is the most used crustacean in ecotoxicological research in parallel to Daphnia<\/i> sp. and has been proposed for the regulatory testing of potential EDCs in the US, Europe and Japan.Major results were: (i) cDNAs encoding the ecdysteroid receptor (EcR) and the retinoid-X-receptor (RXR), were cloned and sequenced, and subsequent molecular phylogenetic analysis (maximum likelihood and neighbor-joining) revealed that the amino acid sequence of the ligand binding domain (LBD) of N. integer<\/i> EcR (NiEcR) clusters as an outgroup of the Crustacea, while NiRXR-LBD clusters in the Malacostracan clade (bootstrap percentage\u00a0=\u00a075%). (ii) 3D-modeling of ligand binding to NiEcR-LBD demonstrated an incompatibility of the insecticide tebufenozide to fit into the NiEcR-ligand binding pocket. This was in great contrast to ponasterone A (PonA) that is the natural molting hormone in Crustacea and for which efficient docking was demonstrated. In addition, the heterodimerization of NiEcR-LBD with the common shrimp Crangon crangon<\/i> (Linnaeus, 1758) RXR-LBD (CrcRXR-LBD) was also modeled in silico<\/i>. (iii) With use of insect Hi5 cells, chimeric constructs of NiEcR-LBD and CrcRXR-LBD fused to either the yeast Gal4-DNA binding domain (DBD) or Gal4-activation domain (AD) were cloned into expression plasmids and co-transfected with a Gal4 reporter to quantify the protein\u2013protein interactions of NiEcR-LBD with CrcRXR-LBD. Investigation of the ligand effect of PonA and tebufenozide revealed that only the presence of PonA could induce dimerization of this heterologous receptor complex. (iv) Finally, in an in vivo<\/i> toxicity assay, N. integer<\/i> juveniles were exposed to tebufenozide at a concentration of 100\u00a0\u00b5g\/L, and no effects against the molting process and nymphal development were scored.In conclusion, the in vitro<\/i> cell reporter assay, based on NiEcR-LBD\/CrcRXR-LBD heterodimerization in Hi5 cells and validated with the natural ecdysteroid hormone PonA, represents a useful tool for the screening of putative EDCs. As a test example for non-steroidal ecdysone agonist insecticides, tebufenozide had no negative effects on NiEcR\/RXR receptor dimerization in vitro<\/i>, nor on the molting process and nymphal development of N. integer<\/i> at the tested concentration (100\u00a0\u00b5g\/L) in vivo<\/i>.","AbstractOtherLang":null,"BibLvlCode":"AS","StandardTitle":"Cloning and functional analysis of the ecdysteroid receptor complex in the opossum shrimp Neomysis integer<\/i> (Leach, 1814)","OrigTitleLangCode":"en","OrigTitleLangCodeExtended":"eng","OrigTitleLangID":15,"DateLastModified":{"date":"2026-04-16 01:31:43.728644","timezone_type":1,"timezone":"+02:00"},"UserAccessRight":null,"UserAccID":null,"AuthorKeywords":"Endocrine-disrupting chemical; Nuclear hormone receptor; Ecdysteroid receptor; Retinoid-X-receptor; Heterologous expression; Heterodimerization; Gal4; Protein modeling; Shrimp exposure assay; Ecdysone agonist; Tebufenozide","OtherDescriptors":null,"Notes":null,"AnaPub":2013,"MonPub":null,"DateUpdate":"2016-06-12","DateCreate":"2013-05-17","SecASFANote":null,"ConfID":null,"PeerRev":1,"VlizCoreFlag":1,"WoScode":"WOS:000317701600005","VABBcode":null,"OpenAcc":0,"DOI":"10.1016\/j.aquatox.2012.12.011"},"refs":null,"anarec":{"AnaID":225700,"PubliDate":2013,"Pagination":"31-40","XtraPublOfAnaID":null,"ISBN":null,"Volume":"130-131","Issue":null,"BRefMon":null,"BRefMonRR":null,"BRefXtra":null,"BRefXtraRR":null,"SerBRefID":42203,"SerRR":"Aquatic Toxicology. 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