title: A novel report of metallothioneins in fish bile: SDS-PAGE analysis, spectrophotometry quantification and metal speciation characterization by liquid chromatography coupled to ICP-MS.
abstract: Metallothioneins (MT) are a validated biomarker for metal exposure. In fish they are usually determined in the liver, while some studies have observed MT in kidney, muscle and gills. There are, however, no reports on MT in fish bile. This study aimed to verify if these proteins are present in fish bile, if they can be used as biomarkers for metal exposure and to characterize the metal speciation present in the different species. Three independent methods to verify MT presence in fish bile were conducted: 1D/2D gel electrophoresis, spectrophotometric analyses and SEC-HPLC-ICP-MS. Results show that all methods verified the presence of these proteins in fish bile, while SEC-HPLC-ICP-MS also characterized also characterized metals responsible for the contamination of the studied sites. Quantification was successfully conducted by two methods, densitometry (1D electrophoresis) and spectrophotometry, and the data indicates that bile MTs follow the same trend as in liver. Therefore, Biliary MT show the potential to be used the same way as MT from other matrices in metal exposure studies, with the advantage of being a simpler matrix and showing the possibility of not sacrificing the animals, as chronic duct cannulation is available.
authors.affiliation: Pontifícia Universidade Católica-Rio de Janeiro-PUC-Rio, Chemistry Department, Bioanalytics Laboratory, Rua Marquês de São Vicente 225, Gávea, CEP: 22453-900, Rio de Janeiro, RJ, Brazil. rachel.hauser.davis@gmail.com
authors.forename: Rachel Ann
authors.initials: RA
authors.lastname: Hauser-Davis
authors.forename: Rodrigo Araújo
authors.initials: RA
authors.lastname: Gonçalves
authors.forename: Roberta Lourenço
authors.initials: RL
authors.lastname: Ziolli
authors.forename: Reinaldo Calixto
authors.initials: RC
authors.lastname: de Campos
mesh_terms: D000818:Animals; D001646:Bile / Q000737:chemistry*; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004781:Environmental Exposure; D005399:Fishes / Q000502:physiology*; D008099:Liver / Q000378:metabolism; D013058:Mass Spectrometry; D008668:Metallothionein / Q000032:analysis* / Q000378:metabolism; D008670:Metals / Q000032:analysis*; D014874:Water Pollutants, Chemical / Q000032:analysis*
chemical_list: D008670:Metals; D014874:Water Pollutants, Chemical; D008668:Metallothionein