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S. I. Karchner S. W. Kennedy S. Trudeau M. E. Hahn 《Marine environmental research》2000,50(1-5):51-56
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) and related planar halogenated aromatic hydrocarbons (PHAHs) are highly toxic to most vertebrate animals, but there are dramatic species differences in sensitivity, both within and among vertebrate classes. For example, studies in cultured avian hepatocytes have revealed differential sensitivity of birds to PHAHs [Kennedy et al. (1996). Toxicol. Appl. Pharmacol., 141, 214-230]. Differences in the characteristics or expression of the aryl hydrocarbon receptor (AHR) could contribute to these species differences in PHAH responsiveness. To investigate the molecular mechanism of differential PHAH sensitivity, we have begun to characterize the AHR in white leghorn chicken (Gallus gallus), Pekin duck (Anas platyrhynchos), and common tern (Sterna hirundo), as well as an amphibian, mudpuppy (Necturus maculosus). Partial AHR cDNAs encompassing the helix-loop-helix and PAS domains were cloned and sequenced. Comparison of amino acid sequences in this region indicated a high degree of sequence conservation among the bird species (97% amino acid identity). The percent identity between bird sequences and either mouse or mudpuppy was lower (79%); the mudpuppy AHR was 74% identical to the mouse AHR. Phylogenetic analysis of these and other AHR amino acid sequences showed that the bird and mudpuppy AHRs were more closely related to mammalian and fish AHR1 forms than to fish AHR2. Future studies include the in vitro expression and functional characterization of AHRs from these and other non-mammalian vertebrates. 相似文献
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Species differences in sensitivity to related planar halogenated aromatic hydrocarbons (PHAH) add significant uncertainty in assessing the ecological risk to aquatic mammals. To investigate mechanisms of PHAH sensitivity in aquatic mammals, we cloned and sequenced the cDNA of Baikal seal aryl hydrocarbon receptor (AHR), an intracellular protein that initiates PHAH-mediated effects. The Baikal seal AHR cDNA has an open reading frame of 843 amino acid residues with a predicted molecular mass of 94.6 kDa. Comparison of AHR amino acid sequences indicated a high degree of sequence conservation (98%) between Baikal and harbor seals. The high conservation of AHRs between Baikal and harbor seals indicates that these seals express AHR proteins closely related structurally. In our previous report (Kim & Hahn, 2002), the dioxin-binding affinity of the harbor seal AHR was at least as high as that of the AHR from a dioxin-sensitive strain of mice, suggesting that this seal species may be sensitive to PHAH effects. This implies that Baikal seal may also be sensitive to dioxin effects. 相似文献
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Newark Bay (NB) killifish (Fundulus heteroclitus) have been chronically exposed to environmental contaminants that activate the aryl hydrocarbon receptor (AHR) and are tolerant to toxic effects and CYP1A induction provoked by AHR ligands. Resistance to CYP1A induction could be due to an epigenetic mechanism such as DNA methylation. We measured in-ovo CYP1A catalytic activity (ethoxyresorufin-O-deethylase, EROD) in NB and reference site killifish embryos aqueously exposed to various concentrations of the de-methylating agent 5-azacytidine, 5-AC (5, 50 and 500 μ(micro)M) with or without 0.2 μ(micro)g/l of the CYP1A inducer 3,3′,4,4′,5 pentachlorobiphenyl (IUPAC PCB126). Neither PCB126 alone, nor PCB126 plus 5-AC, induced EROD above levels in vehicle treated Newark Bay fish. In reference site fish, the same PCB126 dose provoked a 7.4-fold EROD induction relative to controls. We conclude that Newark Bay killifish are resistant to CYP1A induction by co-planar PCBs during early embryological development and our data suggests that DNA methylation does not play a critical role in resistance to CYP1A induction in this model. 相似文献
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The toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and its related planar halogenated aromatic hydrocarbons (PHAHs) are mediated by the aryl hydrocarbon receptor (AHR). To investigate the potential sensitivity to PHAHs and the evolutional diversity of AHR in aquatic birds, AHR cDNAs were initially cloned and sequenced from the livers of a black-footed albatross (Diomedea nigripes) and a common cormorant (Phalacrocorax carbo). In this study, we report the identification of two distinct AHR paralog genes in these species. The two full-length AHR cDNAs from albatross were highly divergent (33% overall amino acid identity, and 60% identity in the N-terminal half). Phylogenetic analysis showed that one of them belongs to the AHR1 clade and the other one to the AHR2 clade, which has been identified only from fishes, but not yet from mammals and birds. Albatross AHR1 encoded a 861-residue protein with a predicted molecular mass of 96.7 kDa, and in the case of albatross AHR2, 925 amino acids and 100.7 kDa. From cormorant liver, the full-length AHR1 cDNA and the partial AHR2 cDNA were cloned. This result strongly suggests that bird species also possess two distinct AHR genes (AHR1 and AHR2). To our knowledge, this is the first report on the presence of an AHR2-like isoform in bird species as well as in fish. 相似文献
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The biotransformation of xenobiotics by microsomal cytochromes P450 is known to be pivotal in the effects of some compounds, and thought to be so for many. A knowledge of CYP gene diversity and CYP function and regulation in aquatic species is pursued, expecting that it will disclose mechanisms, allow predictions regarding species differences in susceptibility, and provide markers for exposure to xenobiotics. As well, it is hoped that such knowledge will provide clues to CYP endogenous functions, and to the origin and functional significance of CYP gene diversity. The knowledge of CYP in marine and other aquatic species is expanding rapidly. The diversity of CYP genes in non-mammalian vertebrates may approximate that in mammals. At present, cloning studies have identified members of gene families 1 to 4 have been cloned from one or more fish species. Where known, the gene structures of fish CYP genes are like those of mammalian homologues. Only one CYP1A gene has been identified in most fish species examined. Fish CYP1As, including multiple forms from recent divergence in some genera, have structural and catalytic properties more like CYP1A1, but also have properties that are 1A2-like, consistent with fish CYP1As representing the CYP ancestral to both CYP1A1 and CYP1A2. A number of genes cloned from several species have been classified in the 3A subfamily. Fish CYP3As catalyze steroid 6β-hydroxylase, and have other properties consistent with mammalian 3As. Recently identified CYP4 genes classify to novel subfamilies but apparently are homologues of mammalian CYP4 genes, and may act on similar substrates. The greatest diversity of fish CYP genes is in family 2; there are now six fish CYP2 subfamilies known. Four of these are novel subfamilies, although cladistic analysis suggests distinct relationships to mammalian CYP2 subfamilies. Heterologous expression and characterization of some of these CYP have identified similar functions among genes in different subfamilies. For example, fish CYP2Ns and CYP2Ps are related to mammalian CYP2Js, and CYP2P3 and CYP2J2 have strikingly similar functions as fatty acid epoxygenases and hydroxylases, with nearly identical regio- and enantioselectivity for metabolism of arachidonic acid. In addition to sequence and catalytic similarities, there also are indications that CYP regulation, tissue and cellular localization are similar between fish and mammals. Yet even in cases where orthology is strongly suggested, e.g. CYP1A, there appear to be taxonomic differences in active site structure suggesting potential differences in involvement of CYP1A in toxicity. In contrast to fish, CYP diversity and functions in aquatic invertebrates are poorly known. Investigators have identified novel gene families and subfamilies in crustaceans (CYP2L; CYP45), molluscs (CYP30, CYP10) and sponges (CYP38). CYP4C genes occur in crustaceans, molluscs and echinoderms, and a new subfamily (CYP4Y) in molluscs. The future? There is no doubt that new CYP will continue to be discovered in non-mammalian vertebrates; some (e.g. CYP51) can be predicted confidently. And, there is no doubt that the numbers known in invertebrates will expand greatly. In insects and C. elegans the numbers are very high, and even slime molds have 18 CYP genes. It is virtually certain that CYP genes with unique functions will be discovered. While the knowledge of CYP genes is increasing, knowledge of CYP function and regulation lag well behind. Technical approaches to speed the aquisition of such knowledge are available. The information will be essential to discern the role that CYP play in the disposition and toxicity of xenobiotics, during development as well as in adults. Yet, when such data are in hand, we may have to face the paucity of information on the diversity, function and regulation other enzymes, notably the glutathione S-transferases, glucuronyl transferases and sulfotransferases, in aquatic species. Discerning orthologous relationships among CYP genes, as well as those for phase II enzymes, could highlight gene lineages associated with conserved and endogenous functions. Understanding CYP endogenous functions, as well as their metabolism of xenobiotics, may reveal fully the ways that chemicals cause toxicity. [Support: Sea Grant NA46RG0470-R/P61, EPA R-829890, NIH ES07381]. 相似文献
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Polycyclic aromatic hydrocarbons (PAHs) are contaminants increasing in the environment largely due to burning of fossil fuels. Our previous work identified a synergistic toxicity interaction in zebrafish embryos occurring when PAHs that are agonists for the aryl hydrocarbon receptor (AHR) co-occur with PAHs that are CYP1A inhibitors. This toxicity is mediated by the AHR2, and morpholino knockdown of CYP1A exacerbated toxicity. This study tested two hypotheses: (1) in the absence of functional CYP1A, metabolism of PAHs is shunted towards CYP1B1, which has been shown in mammals to produce more reactive metabolites of PAHs; alternatively, (2) CYP1B1 serves a protective role similar to CYP1A. We used a morpholino approach to knockdown CYP1B1 alone and in co-knockdown with CYP1A to determine whether we could alter deformities caused by synergistic toxicity of PAHs. CYP1B1 knockdown was not different from non-injected controls; nor were CYP1B1+CYP1A co-knockdown deformities different from CYP1A knockdown alone. These data suggest that CYP1B1 is not a significant factor in causing synergistic toxicity of PAHs, nor, in contrast to CYP1A, in providing protection. 相似文献
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The cytochrome P450 1 (CYP1) family has expanded with the addition of the CYP1B and CYP1C subfamilies. We recently identified a new CYP1 subfamily in zebrafish, CYP1D, with a single gene, CYP1D1. Here we examined sequences found in other fish genomes, i.e., stickleback (Gasterosteus aculeatus) and medaka (Oryzias latipes), for similarities among fish CYP1D1 genes. The full-length deduced amino acid sequences for CYP1D1 in these two species averaged about 43% identity to the CYP1As, but nearly 50% when sequence alignment ambiguities were masked. CYP1D1 has seven exons, similar in size and position to the exons in CYP1D1 and CYP1A in zebrafish. However, the intronic distances were substantially smaller in the medaka and stickleback. There also were differing numbers of putative xenobiotic response elements in the CYP1D1 of the various species. Whether the stickleback or medaka genes are inducible by aryl hydrocarbon receptor (AHR) agonists is yet to be determined. 相似文献
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Tracy K. Collier S.Denise Connor Bich-Thuy L. Eberhart Bernadita F. Anulacion Anders Goksoyr Usha Varanasi 《Marine environmental research》1992,34(1-4)
We are currently analyzing hepatic cytochrome P4501A and associated monooxygenase activities in fish sampled in several regional and national monitoring programs, including the National Benthic Surveillance Project of NOAA's Status and Trends Program, damage assessment studies of the Exxon Valdez oil spill, and intensive surveys of specific embayments, such as Puget Sound, Washington. Thus far, apparent contaminant-related increases in the activities of cytochrome P4501A-dependent monooxygenases have been readily measured in most test species. The results presented in this paper show that, for II species of fish, there is excellent concordance between hepatic activities of aryl hydrocarbon hydroxylase (AHH) and ethoxyresorufin-O-deethylase (EROD); moreover, levels of cytochrome P4501A determined by an enzyme-linked immunosorbent assay (ELISA) are also generally concordant with results from catalytic assays. The use of both a catalytic assay and immunoquantitation is recommended, because of the additional quality assurance provided by concurrent use of an immunoquantitation technique, which is desirable in large monitoring programs. 相似文献
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《African Journal of Marine Science》2013,35(2):215-222
Remotely operated vehicle (ROV) video observations were used to document benthic fauna at a hydrocarbon drilling location, at 2 720 m depth, in the poorly studied deep water off northern Egypt. The decapod Chaceon mediterraneus was the most common organism at the site and the only benthic megafaunal invertebrate observed. Three species of fish, Coryphaenoides mediterraneus, Cataetyx laticeps and Bathypterois mediterraneus, were also encountered. These findings confirmed these three species as the deepest-distributed benthic fish in the eastern Mediterranean, and confirmed that the deep Mediterranean, in particular the eastern basin, is one of the world's poorest deep-sea ecosystems in terms of diversity. The behaviour exhibited by the species observed was consistent with their natatory capacity, deduced from their feeding intensity (gut fullness) and diet (mainly suprabenthic prey were consumed), and conservative life strategies adapted to an extremely low energy environment. 相似文献
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Relative potency of PAHs and heterocycles as aryl hydrocarbon receptor agonists in fish 总被引：6，自引：0，他引：6
The relative potency of polycyclic aromatic compounds as aryl hydrocarbon receptor (AhR) agonists in fish was determined using data on CYP1A induction or AhR binding for 74 polycyclic aromatic hydrocarbons (PAHs) and heterocycles in teleost, avian, or mammalian systems from 18 published papers. Each PAH was assigned a fish potency factor relative to the potency of 2,3,7,8-tetrachlorodibenzo-p-dioxin as an AhR agonist. Two and three ring unsubstituted PAHs were generally inactive in fish, avian, and mammalian systems. Benzo[k]fluoranthene and indeno[1,2,3-cd]pyrene were consistently the most potent PAHs, with fish potency factors of 0.001-0.002. Common structural features associated with higher potency PAHs included 4-6 rings containing fluoranthene or phenanthrene structures with an exposed bay region. These results show that PAHs can have similar potency as many dioxin-like PCBs, and AhR mediated toxicity should be considered in assessing the risks of PAHs in fish. 相似文献
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为构建多鳞(Sillago sihama)遗传连锁图谱并鉴定生长等重要经济性状数量性状位点(QTL),实验通过基因分型测序(GBS)技术对163个多鳞个体(2个亲本和161个全同胞家系F1代)进行测序及标记分型,并通过复合区间定位法对该物种的体重、体高、体厚、眼径、体长和背鳍前长6个生长性状进行QTL定位分析。结果显示,多鳞首张高密度遗传连锁图谱全长2 154.803 c M,标记间平均遗传距离0.455 c M,共有4 735个SNP标记分配到24个连锁群。QTL定位分析结果发现在6个生长性状中共检测到20个生长显著相关QTL位点,分布在8个连锁群上,单个QTL的LOD值范围为3.02～4.23,可解释的表型变异范围为0.14%～8.42%。其中,在连锁群LG08聚集了8个生长性状显著相关的QTL。通过对候选QTL区间内的基因进行功能注释,共筛选到了19个潜在生长调控相关基因,包含igf1、igf2、sstr5、sst1a、tgfbr2、gas1、igfals、gfg6、gfg20、bmp7、kdm5c、tti1以及rbm10等。实验获得的遗传标记及相关候选基因是多鳞生长相关性状标记... 相似文献
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Effects of a nuclear power plant thermal discharge on habitat complexity and fish community structure in Ilha Grande Bay, Brazil 总被引：1，自引：0，他引：1
Tatiana Pires Teixeira Leonardo Mitrano Neves Francisco Gerson Araújo 《Marine environmental research》2009,68(4):188-195
Fish communities and habitat structures were evaluated by underwater visual censuses a rocky location impacted by thermal discharge (I) and at two control locations, one in a Sargassum bed (C1) and the other in a rocky shore with higher structural complexity (C2). Habitat indicators and fish communities exhibited significant differences between the impacted and control locations, with the impacted one showing a significant decrease in fish species richness and diversity, as well as a decrease in benthic cover. At the I location, only 13 fish species were described, and the average water temperature was 32 ± 0.4 °C, compared with 44 species at C1 (25.9 ± 0.3 °C) and 33 species at C2 (24.6 ± 0.2 °C). Significant differences in fish communities among locations were found by ANOSIM with Eucinostomus argenteus, Mugil sp. and Haemulon steindachneri typical of location I, while Abudefduf saxatilis, Stegastes fuscus and Malacoctenus delalandi were typical of the control locations. Our study shows that thermal pollution alters benthic cover and influences fish assemblages by altering composition and decreasing richness. 相似文献
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以乌鳢(Ophiocephalus argus)为研究对象,采用不同日粮饲喂,试验W1组饲喂冰鲜鱼,试验W2组饲喂畜禽内脏。12周后,取肠道内容物,利用16S rDNA技术研究投喂不同饲粮的乌鳢肠道生物群落结构差异和微生物多样性,分别获得W1组和W2组乌鳢肠道细菌有效序列51124和39298条。结果分析表明;W1组和W2组物种注释(operational taxonomic unit,OTU)数目分别为2631和4114条,分类地位明确的细菌种类分别隶属5个属和49个属,W1组和W2组乌鳢肠道细菌种类差异很大。W1组乌鳢肠道细菌的优势类群是邻单胞菌属(Plesiomonas)、狭义梭菌属(Clostridium sensu stricto)、鲸蜡菌属(Cetobacterium);W2组乌鳢肠道细菌的优势类群是狭义梭菌属(Clostridium sensu stricto)、邻单胞菌属(Plesiomonas)、Spartobacteria、甲基副球菌属(Methyloparacoccus)、杆菌属(Alsobacter)、Saccharibacteria、分支杆菌属(Mycobacterium)。W2组乌鳢肠道细菌香农指数高于W1组乌鳢肠道细菌,表明前者肠道细菌多样性大于后者。本实验揭示了畜禽内脏饲喂乌鳢肠道细菌的群落种类多样性、OTU丰富度和物种总数均高于冰鲜鱼饲喂乌鳢;W1组肠道有益菌OTU丰富度大于W2组,且W2组还存在一些潜在致病菌。本研究为乌鳢的健康养殖和后续肠道微生物资源的挖掘提供了理论依据。 相似文献
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本文通过qPCR方法,研究了我国重要海水鱼类——牙鲆（Paralichthys olivaceus）雌、雄成体组织以及性腺分化期的脑、性腺中gnrh1、gnrh2和gnrh3的表达水平。结果显示,gnrh1分布于所有检测的组织：在脑、垂体、肾脏和肝脏中高表达,在性腺中表达相对较低;gnrh2则在雌、雄性的肝脏以及雄性的肠、肾脏、眼睛和头肾中;而gnrh3只在雄性的肾脏、眼睛和肌肉中检测到微弱表达。人工诱导的牙鲆雌核发育鱼苗分别用17β-雌二醇（E2）和17α-甲基睾酮（MT）处理,使其分化为雌性或雄性个体表型。在性腺分化期的脑中,gnrh1的表达呈现上升趋势,在全长（total length,TL）4 cm鱼苗中,E2组的表达显著高于MT组（P<0.05）;gnrh2在2 cm TL时E2组显著高于MT组（P<0.05）,随后在E2和对照组的表达水平均出现下降趋势;gnrh3在2 cm TL时E2和对照组的表达均显著高于MT组（P<0.05）,自6 cm TL时MT和E2组的表达开始下降。在性腺中,三种gnrh在性腺分化前2 cm TL表达量都相对较高,随后呈现下降趋势。综上,推测牙鲆gnrh1可能参与了性腺分化的启动、分化过程及性腺发育,gnrh2主要参与了性腺分化的启动,gnrh3主要参与了性腺分化的启动和分化过程。 相似文献
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三疣梭子蟹HMGR基因的克隆及其在蜕皮中的表达分析 总被引：2，自引：1，他引：1
为了研究3-羟基-3-甲基戊二酰辅酶A还原酶(3-hydroxy-3-methyl glutaryl coenzyme A reductase,HMGR)在甲壳动物蜕皮调控中的作用,采用RT-PCR和c DNA末端快速扩增技术(RACE),克隆得到三疣梭子蟹(Portunus trituberculatus)HMGR基因的c DNA序列(Gen Bank登录号:KF280756)。该序列全长2575bp,包括一个53bp的5′端非编码区,一个686bp的3′端非编码区和一个长度为1836bp的开放阅读框,编码611个氨基酸。该氨基酸序列与已公布的美洲海鳌虾HMGR氨基酸序列相比一致性达65%,具有Ⅰ型HMGR保守催化区域、两个HMG-Co A结合基序和两个NADP(H)结合基序。采用实时荧光定量PCR(q RT-PCR)技术,分析三疣梭子蟹HMGR基因的组织差异表达及在蜕皮周期中的表达水平变化,结果表明HMGR基因在三疣梭子蟹大颚器(MO)中的表达量最高,在其它组织中表达量均极低;在三疣梭子蟹蜕皮周期中,大颚器中HMGR基因的表达量自A期至D0亚期升至最高,然后下降,至D4亚期最低。验证了大颚器是三疣梭子蟹合成甲基法尼酯的唯一器官,表明HMGR在三疣梭子蟹蜕皮调控中起着重要作用。 相似文献