Copper accumulation and toxicity in isolated cells from gills and hepatopancreas of the blue crab (Callinectes sapidus)

Abstract

In the present study, we used fresh preparations of mixed-cell populations to evaluate accumulation and toxicity of dissolved copper (1-100 microM) in isolated cells from posterior gills and hepatopancreas of the blue crab (Callinectes sapidus). For both gill and hepatopancreatic cells, significant increases in copper accumulation were observed after exposure to 50 or 100 microM copper. In gill cells, a linear increase in copper accumulation was observed over time. In hepatopancreatic cells, a maximum level of copper accumulation was achieved after 1 h of exposure, remaining unchanged up to 6 h. After 6 h of exposure, copper content in gill cells was 6.6-fold higher than that in hepatopancreatic cells. In both cell types, copper accumulation always followed a linear relationship with copper concentration in the incubation medium. Significant decreases in cell viability were observed after exposure to either 10 microM copper (gill cells) or 100 microM copper (gill and hepatopancreatic cells). Furthermore, an exponential rise to maximum-type relationship was observed between copper accumulation and toxicity in gill cells. Altogether, these findings indicate that the premise behind the biotic ligand model (BLM) approach is verified in isolated cells from posterior gills of the blue crab (i.e., toxicity is driven by copper accumulation in the biotic ligand, the gill cell). Therefore, these cells can be used as a model for the development of an in vitro BLM version for marine conditions. Isolated cells from the hepatopancreas, however, could be used as a model to better understand the mechanism of copper tolerance at a cellular level in crustaceans.

Multiple functions of the crustacean gill: osmotic/ionic regulation, acid-base balance, ammonia excretion, and bioaccumulation of toxic metals

“…The hemolymph protein hemocyanin binds copper and ensures the transport of the remaining copper to tissues occurs in a less toxic form (Rtal et al, 1996; Truchot and Rtal, 1998). Cells dissociated from the posterior gills of Callinectes sapidus (acclimated to dilute seawater) and incubated in 50 or 100 μM copper, show a linear uptake rate of copper with a several fold higher accumulation in gill cells than in hepatopancreas cells (Paganini and Bianchini, 2009). …”

Consideration of the bioavailability of metal/metalloid species in freshwaters: experiences regarding the implementation of biotic ligand model-based approaches in risk assessment frameworks

“…Cross-species application of BLM has been demonstrated (e.g., Casares et al 2012;Schlekat et al 2010). The use of gill cell lines to study silver toxicity in rainbow trout (Zhou et al 2005) and copper effects in blue crab (Paganini and Bianchini 2009) promise a cost-effective in vitro alternative to using live animals.…”

Cytochemical characterization of gill and hepatopancreatic cells of the crab Ucides cordatus (Crustacea, Brachyura) validated by cell metal transport

“…The hepatopancreas of crustaceans, on the other hand, play a role in absorption and nutrient storage, digestive enzyme synthesis, detoxification of xenobiotics and calcium balance (Lyon & simkiss, 1984;Chavez-Crooker et al, 2001;ziLLi et al, 2003;PaGanini & BianChini, 2009;sterLinG et al, 2010). There are four cell types responsible for these functions: 1.…”