Cytological and biochemical response of R1 cells and isolated hepatocytes from rainbow trout ( Oncorhynchus mykiss) to subacute in vitro exposure to disulfoton

Abstract

Ultrastructural, stereological and biochemical alterations in isolated hepatocytes and the permanent fibrocyte-like cell line R1 from rainbow trout (Oncorhynchus mykiss) exposed to 0, 0.2, 2 and 20 mg/l of the phosphorodithioate pesticide disulfoton (Solvirex, O,O-diethyl S-2-ethylthioethyl phosphorodithioate) for up to 5 days were investigated. In both R1 cells and isolated hepatocytes, distinct dose- and time-dependent morphological alterations including diminished amounts of heterochromatin, proliferation of lysosomal elements, dilation and vesiculation of endoplasmic reticulum cisternae, induction of concentric membrane whorls and an increased amount of lipid droplets could be detected at concentrations of > or = 2 mg/l (R1 cells) and > or = 0.2 mg/l disulfoton (hepatocytes). Additional effects in isolated hepatocytes comprised marginalization of heterochromatin, myelin-like structures attached to mitochondrial membranes, formation of ring-shaped mitochondria, proliferation of smooth endoplasmic reticulum, reduction of rough endoplasmic reticulum, induction of ring-shaped Golgi cisternae, glycogen depletion and occurrence of glycogenosomes. Structural changes in isolated hepatocytes could be correlated to suppression of lactate dehydrogenase, glucose-6-phosphate dehydrogenase, alanine aminotransferase, malic enzyme, esterase as well as glutathione S-transferase, but to a stimulation of 7-ethoxycoumarin-O-deethylase and the rate of lipid peroxidation at concentrations > or = 0.01 mg/l disulfoton. Comparison with data from in vivo experiments with rainbow trout indicate the suitability of in vitro techniques for the evaluation of the toxicological potential of a wide range of ecotoxicologically relevant substances.