Abstract
Copper (Cu) is hyperaccumulated in oyster hemocytes and is an essential trace metal indispensable for diverse innate immune functions. However, the roles of Cu in oyster immune defense are still unclear. In this study, Cu exposure enhanced the phagocytosis of zymosan by increasing the number and length of filopodia, as well as mitochondrial ROS (mitoROS) production mainly in granulocytes, followed by semigranulocytes and agranulocytes. The intracellular calcium level increased to promote the phagosome-lysosome fusion after Cu exposure. The enhancement of phagosomal acidification and mitochondrion-phagosome juxtaposition were also found in granulocytes after Cu exposure. These results indicated that Cu could regulate the phagolysosomal system to enhance the antimicrobial ability of oyster hemocytes with the assistance of mitoROS. Furthermore, Cu(I) and Cu(II) were predominately located in lysosomes, and degranulation may provide a mechanism for exposing Cu to bacteria to prevent their survival and proliferation. Specifically, we showed that the newly formed Cu(I) arising from lysosomal Cu(II) moved to lysosomes and mitochondria in activated hemocytes to induce strong immune responses. The ability of the transformation of Cu(I) from Cu(II) followed granulocytes > semigranlocytes > agranulocytes, indicating that granulocytes played important roles in immune functions of oysters. Our results provided new insights into the understanding of antimicrobial effects of Cu in oyster hemocytes.