Novel Insights into the Role of Copper in Critical Life Stages of Oysters Revealed by High-Resolution NanoSIMS Imaging

Copper (Cu) is a key trace element for many biological processes, but there is little available information regarding its requirements and functions in critical life stages of marine bivalves, during which dramatic morphological and physiological changes occur. In this study, the ontogeny pattern of Cu accumulation across the life history of oysters was explored for the first time and the distributions of Cu in oysters at critical life stages (pediveliger and early settled spat) were in situ mapped by nanoscale secondary ion mass spectrometry (NanoSIMS) with high lateral resolution. We first demonstrated that the late pelagic stage to early settled stage was the critical stage requiring Cu during oyster development. NanoSIMS imaging revealed a significant elevation of intercellular Cu levels along with mitochondrial calcium overload and obvious structural degradation of mitochondria in velar cells at the pediveliger stage, implying the possible role of Cu in cell apoptosis of the velum during larval metamorphosis. Furthermore, an obvious enrichment of Cu together with calcium was observed in the nucleus and mitochondria of ciliated cells of gill tissue at the early settled stage. Their accumulation in the gill cells was significantly higher than that in the juveniles, indicating the potential role of Cu in sustaining the fast growth of the gill filament and the concomitant acceleration of energy metabolism during early benthic development. Our findings offer new insights into the understanding of the interactions between trace metals and marine bivalves.