Accumulation of different metals in oyster Crassostrea gigas: Significance and specificity of SLC39A (ZIP) and SLC30A (ZnT) gene families and polymorphism variation.

The Zrt/Irt-like proteins (ZIP, SLC39A) and zinc transporters (ZnT, SLC30A) are the two major gene families responsible for the import/export of Zn and other metals. In this study, the mRNA expression levels and genetic variations of eight ZnTs and 14 ZIPs were identified in Crassostrea gigas after exposure to Zn, Cd, Cu, Hg, and Pb. Metal exposure induced reactive oxygen species (ROS) and malondialdehyde (MDA) accumulation and antioxidant enzyme expression. The expanded gene numbers of superoxide dismutase (SOD) in the oysters exhibited diverse expression under exposure to the five metals, and the contrasting expressions of both ZnTs and ZIPs under different metal exposures were observed, revealing their ion-specific responses. Zn and Cu have similar transporters and induce high expression levels of ZnT1, 2, 7, and 9 and ZIP1, 3, 6, 9, 10, 11, and 14. Pb induced high expression levels of ZIP7, and 13 and ZnT5, 6, and 7, which are mainly expressed in the endoplasmic reticulum (ER). Cd induced high expression levels of ZnT1, 2, and 7 and ZIP1, 6, 9, 10, 11, and 13. Hg exposure was found to have little effect on the ZIP and ZnT expression levels. Based on 3784 single nucleotide polymorphisms (SNPs) within the ZnTs and ZIPs, genetic association analysis for Zn accumulation was conducted on 427 oyster samples. The 38 SNPs, which were located within 12 genes, were identified to be associated with Zn content (p < 0.01), explaining the phenotypic variation from 1.61% to 3.37%. One nonsynonymous mutation and related haplotypes were identified within ZIP1, explaining 1.69% of the variation in Zn. Its high expression under Zn exposure revealed its important role in Zn transportation. To the best of our knowledge, this study is the first comprehensive investigation of the transportation mechanisms of ZIPs and ZnTs under different metal exposures and the genetic effect of Zn accumulation in oysters, and provides valuable biomarkers and genetic resources to evaluate environmental metal pollution.