Manganese effects on haematopoietic cells and circulating coelomocytes of Asterias rubens (Linnaeus).

Manganese (Mn) is a naturally abundant metal in marine sediments where it mainly occurs as MnO2.During hypoxic conditions it is converted into a bioavailable state, Mn2+, and can reach levels that previouslyhave shown effects on immune competent cells of the crustacean, Nephrops norvegicus. Herewe investigated if Mn also affects circulating coelomocytes and their renewal in the common sea star,Asterias rubens, when exposed to concentrations of Mn that can be found in nature. When the sea starswere exposed to Mn it accumulated in the coelomic fluid and the number of circulating coelomocytes,in contrast to what was recorded in Nephrops, increased significantly. By using the substitute nucleotide,5-bromo-2-deoxyuridine, BrdU, for tracing cell division and by recording mitotic index by nuclei staining,we found that Mn induced proliferation of cells from a putative haematopoietic tissue, the coelomicepithelium. In addition, the haematopoietic tissue and coelomocytes showed stress response in terms ofchanges in HSP70 levels and protein carbonyls, as judged by immunohistochemistry andWestern blotting.Measurement of dehydrogenase activity, using MTS/PMS, revealed that Mn showed cytotoxic properties.We also found that the phagocytotic capacity of coelomocytes was significantly inhibited by Mn. It wasconcluded that the exposure of A. rubens to Mn induced renewal of coelomocytes and impaired theirimmune response.