METAL REGULATION AND MOLTING IN THE BLUE CRAB, CALLINECTES SAPIDUS: METALLOTHIONEIN FUNCTION IN METAL METABOLISM.

We recently demonstrated that zinc, copper, and hemocyanin metabolism in the blue crab varies as a function of the molt cycle. To extend these observations, and better delineate metal metabolism in marine crustaceans, we have conducted experiments to determine if environmental temperature and season of the year affect concentrations of hemocyanin and copper in the hemolymph and copper and zinc in the digestive gland. Overwintering, cold water crabs (6°C) had decreased hemocyanin and copper in the hemolymph and normal zinc and copper in the digestive gland with respect to summer crabs collected at 20-30°C. When these crabs were warmed to 20°C and fed fish for three weeks, they showed increases in the concentrations of copper in the digestive gland, and copper and hemocyanin in the hemolymph. In addition, a change from a zinc to a copper-dominated metallothionein was found in a majority of the warmed crabs, suggesting the involvement of copper metallothionein in the resynthesis of hemocyanin. Based on these observations and previous data (Engel, 1987) a conceptual model of copper and zinc partitioning in the blue crab has been constructed. In this model, metallothionein has an important role in metal regulation both during molting and in the changes related to season of the year. Metallothionein-bound copper and zinc appear to be regulated at the cellular level for the synthesis of metalloproteins, such as hemocyanin (copper) and carbonic anhydrase (zinc), both of which are necessary for normal growth and survival. Finally, we present evidence showing that copper metallothionein can directly transfer its metal to the active site of apohemocyanin. Copper insertion seems to precede the formation of viable oxygen binding sites.