title: Mechanistic toxicodynamic model for receptor-mediated toxicity of diazoxon, the active metabolite of diazinon, in Daphnia magna.
abstract: The organothiophosphate diazinon inhibits the target site acetylcholinesterase only after activation to its metabolite diazoxon. Commonly, the toxicity of xenobiotics toward aquatic organisms is expressed as a function of the external concentration and the resulting effect on the individual level after fixed exposure times. This approach does not account for the time dependency of internal processes such as uptake, metabolism, and interaction of the toxicant with the target site. Here, we develop a mechanistic toxicodynamic model for Daphnia magna and diazoxon, which accounts for the inhibition of the internal target site acetylcholinesterase and its link to the observable effect, immobilization, and mortality. The model was parametrized by experiments performed in vitro with the active metabolite diazoxon on enzyme extracts and in vivo with the parent compound diazinon. The mechanism of acetylcholinesterase inhibition was shown to occur irreversibly in two steps via formation of a reversible enzyme-inhibitor complex. The corresponding kinetic parameters revealed a very high sensitivity of acetylcholinesterase from D. magna toward diazoxon, which corresponds well with the high toxicity of diazinon toward this species. Recovery of enzyme activity but no recovery from immobilization was observed after in vivo exposure to diazinon. The toxicodynamic model combining all in vitro and in vivo parameters was successfully applied to describe the time course of immobilization in dependence of acetylcholinesterase activity during exposure to diazinon. The threshold value for enzyme activity below which immobilization set in amounted to 40% of the control activity. Furthermore, the model enabled the prediction of the time-dependent diazoxon concentration directly present at the target site.
authors.affiliation: Eawag, Swiss Federal Institute of Aquatic Science and Technology, 8600 Dübendorf, Switzerland.
authors.forename: Andreas
authors.initials: A
authors.lastname: Kretschmann
authors.forename: Roman
authors.initials: R
authors.lastname: Ashauer
authors.forename: Kristina
authors.initials: K
authors.lastname: Hitzfeld
authors.forename: Piet
authors.initials: P
authors.lastname: Spaak
authors.forename: Juliane
authors.initials: J
authors.lastname: Hollender
authors.forename: Beate I
authors.initials: BI
authors.lastname: Escher
mesh_terms: D000818:Animals; D002800:Cholinesterase Inhibitors / Q000633:toxicity*; D003621:Daphnia / Q000187:drug effects* / Q000201:enzymology; D003976:Diazinon / Q000378:metabolism / Q000633:toxicity; D007700:Kinetics; D008954:Models, Biological; D009943:Organophosphorus Compounds / Q000633:toxicity*
chemical_list: D002800:Cholinesterase Inhibitors; D009943:Organophosphorus Compounds; C000912:diazoxon; D003976:Diazinon