Institute of Aquaculture, University of Stirling, UK.
Comp Biochem Physiol C Toxicol Pharmacol. 2011 Aug;154(2):93-9. doi: 10.1016/j.cbpc.2011.03.010. Epub 2011 Apr 4.
Copper transporting ATPase, ATP7A, is an ATP dependent copper pump present in all vertebrates, critical for the maintenance of intracellular and whole body copper homeostasis. Effects of copper treatment on ATP7A gene expression in fibroblast cells (SAF1) of the sea bream (Sparus aurata) were investigated by qRT-PCR and by a medium density microarray from a closely related species, striped sea bream (Lithognathus mormyrus). To discriminate between the effects of Cu and other metals, SAF1 cells were exposed to sub-toxic levels of Cu, Zn and Cd. Expression of Cu homeostasis genes copper transporter 1 (CTR1), Cu ATPase (ATP7A), Cu chaperone (ATOX1) and metallothionein (MT) together with the oxidative stress markers glutathione reductase (GR) and Cu/Zn superoxide dismutase (CuZn/SOD) were measured 0, 4 and 24 hours post-exposure by qRT-PCR. Microarray was conducted on samples from 4 hours post Cu exposure. Cu, Zn and Cd increased MT and GR mRNA levels, while only Cu increased ATP7A mRNA levels. Microarray results confirmed the effects of Cu on ATP7A and MT and in addition showed changes in the expression of genes involved in protein transport and secretion. Results suggest that ATP7A may be regulated at the transcriptional level directly by Cu and by a mechanism that is different from that exerteted by metals on MT genes.
铜转运 ATP 酶（ATP7A）是一种依赖于 ATP 的铜泵，存在于所有脊椎动物中，对维持细胞内和全身铜稳态至关重要。通过 qRT-PCR 和来自密切相关物种条纹石鲷（Lithognathus mormyrus）的中密度微阵列，研究了铜处理对真鲷（Sparus aurata）成纤维细胞（SAF1）中 ATP7A 基因表达的影响。为了区分 Cu 和其他金属的影响，将 SAF1 细胞暴露于亚毒性水平的 Cu、Zn 和 Cd 下。通过 qRT-PCR 在暴露后 0、4 和 24 小时测量铜稳态基因铜转运蛋白 1（CTR1）、Cu ATP 酶（ATP7A）、Cu 伴侣（ATOX1）和金属硫蛋白（MT）以及氧化应激标志物谷胱甘肽还原酶（GR）和 Cu/Zn 超氧化物歧化酶（CuZn/SOD）的表达。在 Cu 暴露后 4 小时进行微阵列。Cu、Zn 和 Cd 增加了 MT 和 GR mRNA 水平，而只有 Cu 增加了 ATP7A mRNA 水平。微阵列结果证实了 Cu 对 ATP7A 和 MT 的影响，并显示了参与蛋白质运输和分泌的基因表达的变化。结果表明，ATP7A 可能直接受 Cu 的转录调控，并且其机制与金属对 MT 基因的作用不同。
