Understanding copper sensitivity in zebrafish (Danio rerio) through the intracellular localization of copper transporters in a hepatocyte cell-line ZFL and the tissue expression profiles of copper transporters

Zebrafish (Danio rerio) is a freshwater fish species of Cyprinidae known for its copper (Cu) intolerance, yet the underlying mechanisms of the sensitivity remain unclear. In this study, we examined the highly conserved molecular machineries in the copper handling system, namely ATOX1, ATP7A, ATP7B, and CTR1, by profiling their gene expression patterns among tissues before and after acute waterborne Cu exposure, and investigating their intracellular localization patterns using a zebrafish hepatocyte cell line, ZFL. We found that ATP7B was weak in its response toward Cu exposure to elicit its copper efflux function. Tissue distribution of these Cu transporters, however, revealed a distinct expression profile compared with mammals and other fish, particularly ATP7A, which unlike ATP7B was highly expressed in the liver, while ATP7B, not ATP7A, was specifically expressed in the intestine. ATOX1 transcript expression was also found to be significantly up-regulated with acute waterborne Cu, in contrast to the decreased expression found in other fish. A possible explanation for the Cu sensitivity in zebrafish is discussed.