Metallothionein-like proteins in the freshwater oligochaete Limnodrilus udekemianus and their role as a homeostatic mechanism against cadmium toxicity

Abstract

The purpose of this study was to determine if metallothioneins are present in the aquatic oligochaete Limnodrilus udekemianus and to determine the interplay between the presence of these proteins, cadmium (Cd) exposure, and Cd toxicity. The latter was geared specifically towards evaluating the role of metallothionein as a homeostatic mechanism against Cd toxicity. These issues are important for evaluating the usefulness of the quantification of metallothioneins as a biomonitoring tool. Worms in sediment were exposed to Cd under static conditions, with Cd initially added to the aqueous phase. Survival was monitored while respiration (as a measure of sublethal Cd effects) was determined immediately following exposure. Metallothioneins were separated from the cytosol by gel permeation high performance liquid chromatography (HPLC) while Cd levels were quantified in whole worms, cytosol and cytosolic fractions. Also, a Cd-saturation assay was used to determine the amounts of Cd bound to metallothionein and the total Cd-binding capacity of the metallothionein. Limnodrilus udekemianus has a metallothionein-like protein (an inducible cytosolic protein with an apparent molecular weight of approximately 15 kD that binds high levels of Cd and shows a red shift upon Cd binding). Sediment Cd levels above 60 μg/g were lethal to the worms (in 8-day exposures). Respiration rates at 13 and 41 μg/g Cd were not significantly different from controls, though cytosolic Cd levels were substantially increased in the 41 μg/g exposure. In this latter cytosol, Cd levels were significantly elevated in the low molecular weight pool (which includes metallothioneins) but not in the other pools, while the Cd-saturation assay also showed that worms in this group had significantly elevated levels of metallothionein-bound Cd. However, in all treatments the metallothionein was far from saturated by Cd. These observations indicate that no ‘spill-over’ of Cd was evident as lethal levels of Cd were approached. The overall cytosolic Cd distribution, and the degree of metallothionein saturation in Limnodrilus udekemianus thus do not appear to be good predictors of Cd toxicity in this species.