Occurrence and distribution of antioxidant enzymes in the haemolymph of the shore crab Carcinus maenas

The haemocytes and cell-free haemolymph of the shore crab Carcinus maenas were examined for the presence of the antioxidant enzymes, catalase (EC 1.11.1.6), glutathione peroxidase (GPX) (EC 1.11.1.9) and superoxide dismutase (SOD) (EC 1.15.1.1) by biochemical assay and immunocytochemistry. The results show that all three enzymes are present in the haemocytes, but vary in distribution and amount. Catalase was detected principally in the phagocytic hyaline cells, whereas GPX and SOD were found in both the hyaline and granular cells. Catalase and glutathione peroxidase activities were absent from the semi-granular cells, and although SOD was detected in some samples, there was enormous variation in the level of this enzyme between individuals. Cell-free haemolymph, by contrast, was found to be positive for GPX and SOD but lacked catalase activity. Immunocytochemical analyses further showed that only 2 to 16% of the cells stain positively for these enzymes, and where present the enzymes tended to be distributed throughout the cytoplasm. With the granular cells, however, there was some localization of GPX and SOD around the granules. Since the phagocytes of C. maenas are known to produce superoxide anion radicals, hydrogen peroxide, and possible other reactive oxygen species upon stimulation with phorbol myristate acetate, lipopolysaccharide, or concanavalin A in vitro, it is likely that catalase, GPX and SOD are present in the haemolymph to assist in protection of the host tissues against oxyradical damage during host defence in vivo.