Contaminant impact on interactions of molecular probes with lysosomes in living hepatocytes from dab Limanda limanda

Abstract

During the Bremerhaven Workshop, effects of contaminants along a well characterised North Sea contaminant gradient heading northwest of the port of Bremerhaven in the German Bight were determined in dab Limanda lirnanda using i n vitro techniques on isolated hepatocytes. Results demonstrated that lysosomal membrane integrity was impaired following contaminant exposure which resulted in an inability to retain the supravital dye neutral red and in reduced lysosomal uptake of the fluorescent probe acridine orange. There was an improvement in the condition of exposed hepatocytes in samples taken further offshore, and away from the source of the contaminant, however, this trend was reversed in samples taken from the furthermost station on the northeast tail of the Dogger Bank. These findings are related to the histopathological evidence of abnormal lysosomal enlargement and disturbance of liver cell structure.

Toxicological pathology of dab Limanda limanda along pollution gradients in the southern North Sea

“…The presence of the high levels of neutral lipid in livers of dab from Stn 3 coupled with the other hepatic changes described would suggest a toxic effect, such as fatty change, rather than a physiological over-production of normal storage lipid. Over-all, the changes reported here are consistent with a toxic effect in the liver of dab from Stn 3 and are strengthened by the presence of lysosome impairment, reduced endocytosis, increased low density lipoprotein, increased oxyradical formation, increased endoplasmic reticulum, elevated EROD, elevated P4501A1 and the detection of ras-oncoprotein, also in livers of dab from Stn 3 (Kohler et al 1992, Lowe et al 1992, Moore 1992a, b, Moore & Evans 1992, Renton & Addison 1992. In total the data from these varied biomarkers suggest toxic liver injury in dab taken from the inshore stations.…”

“…These pathological methods of investigation, which were mainly limited to gross inspection, have revealed a variety of spatial and temporal trends in fish disease patterns in the North Sea (Bucke et al 1984, Bucke 1988, 1991, Bucke & Waterman 1988, Dethlefson 1988. This epidemiological type of approach can be complemented by the application of techniques which operate at the molecular, cellular and tissue level of organisation (Kohler 1989, Moore 1990, 1992a, b, Moore & Simpson 1991, Stegeman & Lech 1991, Varanasi & Stein 1991, Cameron & Berg 1992, Kohler et al 1992, Lowe et al 1992, Moore & Evans 1992. The combined methodology coupled with the use of supportive laboratory bioassays should help to define causal links between some classes of contaminant and specific types of toxic injury in fish.…”

In vitro effect of cadmium and copper on separated blood leukocytes of Dicentrarchus labrax

“…The cytotoxic effects of cadmium and copper are reflected in the alteration of the lysosomal membrane, probably due to the high levels of metals that may accumulate in lysosomes rather than being eliminated from the body (Moore, 1990;Regoli, 1992). It is likely that Cd and Cu lead to the inhibition of Mg 2 þ -ATPase, a proton pump of lysosomal membranes that maintains the internal environment of lysosomes acid (Lowe et al, 1992). Dysfunction of this ATPase allows the free passage of lysosomal contents, including NRU, into the cytosol (Lowe et al, 1995).…”

Biomarkers responses of the earthworm Eisenia fetida to acetochlor exposure in OECD soil

“…One method of determining lysosomal membrane stability is by using a neutral red retention assay. This technique has already proved to be reliable, dose-related and practical to be used in marine (Lowe et al, 1992) and terrestrial systems (Weeks and Svendsen, 1996). The assay has shown to be sensitive to various heavy metals and pesticide pollution (Lowe et al, 1992;Eason et al, 1999;Robidoux et al, 2002).…”