Metallothionein induction by Cu, Cd and Hg in Dicentrarchus labrax liver: assessment by RP-HPLC with fluorescence detection and spectrophotometry.
Metallothionein was quantified in sea bass Dicentrarchus labrax intraperitoneally (i.p.) injected with different Cu, Cd and Hg doses (50-250 microg kg(-1) wet wt) after 48 h exposure. A distinct peak with 16.8 min retention time was obtained by reversed-phase high performance liquid chromatography coupled to fluorescence detection (RP-HPLC-FD) with the three metals. Total metallothionein levels assayed in unheated liver extracts by RP-HPLC-FD were significantly higher (1.3-1.95-fold) than those obtained by the well-established spectrophotometric method. In the RP-HPLC-FD method, metallothionein increased linearly with Cu and Hg doses, being saturated beyond 100 mug kg(-1) Cd. Maximum induction was obtained at 100 microg kg(-1) Cd (5.3-fold), and 250 microg kg(-1) Cu or Hg (8- and 5.1-fold, respectively). At low doses no metallothionein induction was shown by the less sensitive spectrophotometric assay.
Jebali
Jamel
J
Laboratoire de Biochimie et de Toxicologie Environmentale, Institut Supérieur d'Agronomie de Chott-Mariem, 4042 Sousse, Tunisia.
Banni
Mohamed
M
Gerbej
Hamadi
H
Boussetta
Hamadi
H
López-Barea
Juan
J
Alhama
José
J
D000818:Animals; D001492:Bass / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D008099:Liver / Q000187:drug effects* / Q000378:metabolism; D008668:Metallothionein / Q000096:biosynthesis* / Q000235:genetics; D019216:Metals, Heavy / Q000633:toxicity*; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D015854:Up-Regulation / Q000187:drug effects*; D014874:Water Pollutants, Chemical / Q000633:toxicity*
D019216:Metals, Heavy; D014874:Water Pollutants, Chemical; D008668:Metallothionein