Dept. of Biological Sciences, Oakland University, Rochester, MI 48309, USA.
BMC Mol Biol. 2009 Nov 25;10:104. doi: 10.1186/1471-2199-10-104.
Arsenic is one of the most ubiquitous toxins and endangers the health of tens of millions of humans worldwide. It is a mainly a water-borne contaminant. Inorganic trivalent arsenic (AsIII) is one of the major species that exists environmentally. The transport of AsIII has been studied in microbes, plants and mammals. Members of the aquaglyceroporin family have been shown to actively conduct AsIII and its organic metabolite, monomethylarsenite (MAsIII). However, the transport of AsIII and MAsIII in in any fish species has not been characterized.
In this study, five members of the aquaglyceroporin family from zebrafish (Danio rerio) were cloned, and their ability to transport water, glycerol, and trivalent arsenicals (AsIII and MAsIII) and antimonite (SbIII) was investigated. Genes for at least seven aquaglyceroporins have been annotated in the zebrafish genome project. Here, five genes which are close homologues to human AQP3, AQP9 and AQP10 were cloned from a zebrafish cDNA preparation. These genes were named aqp3, aqp3l, aqp9a, aqp9b and aqp10 according to their similarities to the corresponding human AQPs. Expression of aqp9a, aqp9b, aqp3, aqp3l and aqp10 in multiple zebrafish organs were examined by RT-PCR. Our results demonstrated that these aquaglyceroporins exhibited different tissue expression. They are all detected in more than one tissue. The ability of these five aquaglyceroporins to transport water, glycerol and the metalloids arsenic and antimony was examined following expression in oocytes from Xenopus leavis. Each of these channels showed substantial glycerol transport at equivalent rates. These aquaglyceroporins also facilitate uptake of inorganic AsIII, MAsIII and SbIII. Arsenic accumulation in fish larvae and in different tissues from adult zebrafish was studied following short-term arsenic exposure. The results showed that liver is the major organ of arsenic accumulation; other tissues such as gill, eye, heart, intestine muscle and skin also exhibited significant ability to accumulate arsenic. The zebrafish larvae also accumulate considerable amounts of arsenic.
This is the first molecular identification of fish arsenite transport systems and we propose that the extensive expression of the fish aquaglyceroporins and their ability to transport metalloids suggests that aquaglyceroporins are the major pathways for arsenic accumulation in a variety of zebrafish tissues. Uptake is one important step of arsenic metabolism. Our results will contribute to a new understanding of aquatic arsenic metabolism and will support the use of zebrafish as a new model system to study arsenic associated human diseases.
砷是最普遍存在的毒素之一,危害着全球数千万人的健康。它主要是一种水污染物。无机三价砷(AsIII)是环境中存在的主要物种之一。已经研究了微生物、植物和哺乳动物中 AsIII 的运输。水通道甘油蛋白家族的成员已被证明能够主动传导 AsIII 及其有机代谢物一甲基砷酸盐(MAsIII)。然而,在任何鱼类物种中,AsIII 和 MAsIII 的运输都没有被描述过。
在这项研究中,从斑马鱼(Danio rerio)中克隆了水通道甘油蛋白家族的五个成员,并研究了它们运输水、甘油以及三价砷(AsIII 和 MAsIII)和锑(SbIII)的能力。斑马鱼基因组项目中至少有七个水通道甘油蛋白基因被注释。在这里,从斑马鱼 cDNA 制备物中克隆了五个与人类 AQP3、AQP9 和 AQP10 密切同源的基因。根据与相应的人类 AQPs 的相似性,这些基因被命名为 aqp3、aqp3l、aqp9a、aqp9b 和 aqp10。通过 RT-PCR 检测了 aqp9a、aqp9b、aqp3、aqp3l 和 aqp10 在多种斑马鱼器官中的表达。我们的结果表明,这些水通道甘油蛋白表现出不同的组织表达。它们都在不止一种组织中被检测到。在非洲爪蟾卵母细胞中表达这些五个水通道甘油蛋白后,检测了它们运输水、甘油以及金属砷和锑的能力。这些通道都以相当的速率显示出大量的甘油转运。这些水通道甘油蛋白还促进了无机 AsIII、MAsIII 和 SbIII 的摄取。在短期砷暴露后,研究了鱼类幼虫和成年斑马鱼不同组织中的砷积累。结果表明,肝脏是砷积累的主要器官;鳃、眼、心脏、肠肌和皮肤等其他组织也表现出显著的砷积累能力。斑马鱼幼虫也积累了相当数量的砷。
这是首次对鱼类亚砷酸盐转运系统进行分子鉴定,我们提出鱼类水通道甘油蛋白的广泛表达及其转运金属的能力表明,水通道甘油蛋白是砷在各种斑马鱼组织中积累的主要途径。摄取是砷代谢的重要步骤。我们的结果将有助于对水生砷代谢的新认识,并支持将斑马鱼作为研究与砷相关的人类疾病的新模型系统。