Zinc is incorporated into the exoskeleton during post-ecdysial mineralization and inhibits exoskeletal calcification in the blue crab, Callinectes sapidus

Abstract

Zinc is an essential but toxic metal with both natural and anthropogenic sources. Zinc has been reported to be present in crustacean exoskeleton, but it has remained unknown as to when zinc is incorporated into the shell during the molting cycle and the effects zinc has on exoskeleton properties. This study was conducted to identify a period during the molting cycle, in which zinc is incorporated into the exoskeleton of the blue crab, Callinectes sapidus, and to identify deleterious effects of zinc incorporation on the exoskeleton. It was hypothesized that zinc would be incorporated into the shell during the mineralization phase using calcium transporters, which would inhibit exoskeletal calcification. Post-ecdysial blue crabs were given two injections of zinc in the form of zinc chloride dissolved in Pantin's crustacean saline at the arthrodial membrane at three treatment levels: 0.0, 1.0 and 5.0 µg Zn/g wet weight. Exoskeletal and hemolymph samples were then analyzed for zinc, calcium, and magnesium content. Gill, muscle, and hepatopancreas samples were analyzed for zinc only. Epidermis samples were analyzed for carbonic anhydrase activity. The results showed that the injection dose of 1.0 µg Zn/g wet weight resulted in significant accumulation of zinc in the exoskeleton. There was no significant accumulation of exoskeletal zinc following 5.0 µg Zn/g wet weight injections. A significant reduction in exoskeletal calcium content in crabs treated with 1.0 or 5.0 µg Zn/g wet weight was also observed. The hypothetical model explaining zinc's incorporation into the exoskeleton and inhibition of exoskeletal calcification is proposed. Additionally, for the soft tissues examined, significant zinc accumulation was only observed in the hepatopancreas following zinc treatment. Our data points to the existence of crustacean zinc transporter. This study is the first to present evidence that zinc is deposited to the exoskeleton during post-ecdysial mineralization and inhibits exoskeletal calcification in a crustacean.