Molecular analysis and heavy metal detoxification of ABCC1/MRP1 in zebrafish
Molecular analysis and heavy metal detoxification of ABCC1/MRP1 in zebrafish
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DOI：
10.1007/s11033-010-0283-z
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发表时间：
2011-03-01
影响因子：
2.8
通讯作者：
Cui, Zongbin
中科院分区：
文献类型：
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作者：
Long, Yong;Li, Qing;Cui, Zongbin
ABCC1/MRP1属于ATP结合盒超家族，其表达升高与多种肿瘤细胞的多药耐药密切相关。在正常组织中，ABCC1 赋予对多种外源物质和毒物的抵抗力，证明了其在组织防御中的重要作用。在这里，我们报告了斑马鱼 abcc1 基因的克隆和功能表征。该基因位于斑马鱼 3 号染色体上，包含 4,557 bp 的开放阅读框。推导的多肽由1,518个氨基酸组成，与人ABCC1有70%的一致性。系统发育分析表明，来自 13 种脊椎动物的 ABCC1 蛋白在进化过程中高度保守。通过整体原位杂交和实时 PCR 检测斑马鱼 abcc1 基因在发育胚胎中的转录表达。在四细胞阶段胚胎中可检测到斑马鱼 abcc1 基因的转录本，表明该基因是母系表达的。 ABCC1 mRNA在受精后12小时前广泛分布于胚胎中，24至72小时主要位于眼睛和大脑，96至120小时主要位于鳃。此外，斑马鱼abcc1基因在1-hpf胚胎中高表达，并在所有检查的成体组织中检测到，其中在睾丸中表达最高，在心脏和肝脏中表达最低。将ZF4细胞和胚胎暴露于CdCl2中心点2.5H(2)O、HgCl2、Pb(NO3)(2)或Na3AsO4中心点12H(2)O显着诱导abcc1基因的转录表达。此外，abcc1的过度表达提高了暴露于Cd、Hg或As的胚胎的存活率，而abcc1突变体(ABCC1-G1420D)的过度表达使斑马鱼胚胎对有毒金属敏感。这些数据表明斑马鱼 ABCC1 在重金属解毒中具有至关重要的作用。
ABCC1/MRP1 belongs to the ATP-binding cassette superfamily and its elevated expression is closely associated with the multidrug resistance of various tumor cells. In normal tissues, ABCC1 confers resistance to a wide variety of xenobiotics and toxicants, demonstrating its important roles in tissue defense. Here, we report the cloning and functional characterization of abcc1 gene in zebrafish. This gene is localized on zebrafish chromosome 3 and contains a 4,557 bp open-reading frame. The deduced polypeptide is composed of 1,518 amino acids, which shares 70% identity with human ABCC1. Phylogenetic analysis revealed that ABCC1 proteins from thirteen vertebrate species are highly conserved during evolution. Transcriptional expression of zebrafish abcc1 gene in developing embryos was examined by whole-mount in situ hybridization and real-time PCR. Transcripts of zebrafish abcc1 gene were detectable in four-cell stage embryos, indicating that this gene is maternally expressed. ABCC1 mRNAs were ubiquitously distributed in embryos before 12 h post-fertilization (hpf) and mainly localized in eyes and brain from 24 to 72 hpf, and in gills from 96 to 120 hpf. In addition, zebrafish abcc1 gene was highly expressed in 1-hpf embryos and detected in all adult tissues examined, with highest expression in testis and lowest in heart and liver. Exposure of ZF4 cells and embryos to CdCl2 center dot 2.5H(2)O, HgCl2, Pb(NO3)(2), or Na3AsO4 center dot 12H(2)O significantly induced transcriptional expression of abcc1 gene. Furthermore, overexpression of abcc1 improved the survival rates of embryos exposed to Cd, Hg or As, while overexpression of a abcc1 mutant (ABCC1-G1420D) sensitized zebrafish embryos to toxic metals. These data indicate that zebrafish ABCC1 has crucial roles in heavy metals detoxification.