Limited species differences in estrogen receptor alpha-medicated reporter gene transactivation by xenoestrogens

Sumida, Kayo; Ooe, Norihisa; Saito, Koichi; Kaneko, Hideo

doi:10.1016/s0960-0760(03)00003-7

Cited by 23 publications

(21 citation statements)

References 29 publications

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“…When 10 pM E 2 was added, HeLa cells exhibited an increase in luciferase activity with human ERα and ERβ, whereas the other ERs required 100 pM to 1 nM E 2 to show such an increase. This corresponds with the previous report of a shift in dose‐response curves for E 2 rainbow trout ERα as compared to human ERα [15]. Petit et al [30] also reported similar findings from gene transactivation experiments.…”

Section: Discussionsupporting

confidence: 91%

“…The control plasmid continuously expresses the luciferase protein, and when a chemical exerts toxic effects, the enzyme activity is lowered. As described in previous reports, we here determined the maximum doses of test chemicals using the results of the control assay (data not shown) [15,17].…”

Section: Discussionmentioning

confidence: 99%

“…Endocrine disruption by chemicals is caused by two main mechanisms: Competitive binding to hormone receptors, and modification of the hormone biosynthesis pathways. We have focused on competitive binding to the estrogen receptors (ERs) of chemicals and have examined species differences (human, rat, chicken, alligator [ Caiman crocodilus ], whiptail lizard [ Cnemidophorus uniparens ], African clawed frog, and rainbow trout) in transactivation by means of reporter gene (luciferase) assays using HeLa cells with full‐length ER cDNAs cloned by reverse‐transcription polymerase chain reaction [15]. No significant variation in transactivation of the reporter gene has been found among human, rat, chicken, alligator, whiptail lizard, and African clawed frog ERs.…”

Section: Introductionmentioning

confidence: 99%

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Molecular cloning of estrogen receptors from fathead minnow (Pimephales promelas) and bluegill (Lepomis macrochirus) fish: Limited piscine variation in estrogen receptor–mediated reporter gene transactivation by xenoestrogens

Sumida

Saito

2008

Environmental Toxicology and Chemistry

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Estrogen receptors (ERs) play essential roles in estrogen function. Because they may be targets of environmental chemicals, concern exists that exposure to some anthropogenic agents may result in disruption of endocrine systems in human and wildlife populations. Various kinds of ERs have been reported, and a number of in vitro assays have been established to test hormone-like activity of chemicals. In the present study, cDNAs encoding ERs from fathead minnow (Pimephales promelas) and bluegill (Lepomis macrochirus) fish, which are useful for environmental monitoring, were cloned and sequenced. Four complete, full-length ERalpha and ERbeta cDNA sequences could be derived, with an ATG start site and a TGA termination signal included in each. Fathead minnow ERalpha has 602 amino acid residues, bluegill ERalpha1 has 582, bluegill ERalpha2 has 506, and bluegill ERbeta has 554. When the cDNAs were inserted into the pRc/RSV vector and transfected into HeLa cells (derived from cervical cancer cells) with a reporter plasmid, the encoding proteins were confirmed to be functional through interaction of the receptor-ligand complex with the estrogen-responsive element (ERE). Here, we report the analysis of piscine differences in ER-dependent transactivation with some chemicals using reporter gene assays. In agonist assays, fold-induction of luciferase activity significantly varied among ERs, but responsiveness to each chemical demonstrated certain similarities. These reporter gene assays appear to be very useful for estimating adverse effects of chemicals, including hormone-like or antihormone-like activity, in fish.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Molecular cloning of estrogen receptors from fathead minnow (Pimephales promelas) and bluegill (Lepomis macrochirus) fish: Limited piscine variation in estrogen receptor–mediated reporter gene transactivation by xenoestrogens

Sumida

Saito

2008

Environmental Toxicology and Chemistry

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“…There is conflicting historical evidence regarding whether the sequence differences among receptors from different classes translate into meaningful functional differences [7][8][9][10]. Significant differences in sensitivity and chemical binding have been observed between hER and rainbow trout estrogen receptor (rtER) [8,9,11,12], as well as between hER and alligator ER [13].…”

Section: Introductionmentioning

confidence: 99%

Comparison of chemical binding to recombinant fathead minnow and human estrogen receptors alpha in whole cell and cell-free binding assays

Rider

Hartig

Cardon

et al. 2009

Environmental Toxicology and Chemistry

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Mammalian receptors and assay systems are generally used for in vitro screening of endocrine-disrupting chemicals with the assumption that minor differences in amino acid sequences among species do not translate into significant differences in receptor function. Objectives of the present study were to evaluate the performance of two different in vitro assay systems (a whole cell and a cell-free competitive binding assay) in assessing whether binding of chemicals differs significantly between full-length recombinant estrogen receptors from fathead minnows (fhERalpha) and those from humans (hERalpha). It was confirmed that 17beta-estradiol displays a reduction in binding to fhERalpha at an elevated temperature (37 degrees C), as has been reported with other piscine estrogen receptors. Several of the chemicals (17beta-estradiol, ethinylestradiol, alpha-zearalanol, fulvestrant, dibutyl phthalate, benzyl butyl phthalate, and cadmium chloride) displayed higher affinity for fhERalpha than for hERalpha in the whole cell assay, while only dibutyl phthalate had a higher affinity for fhERalpha than for hERalpha in the cell-free assay. Both assays were effective in identifying strong binders, weak binders, and nonbinders to the two receptors. However, the cell-free assay provided a less complicated and more efficient binding platform and is, therefore, recommended over the whole cell binding assay. In conclusion, no strong evidence showed species-specific binding among the chemicals tested.

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“…Such systems have been used in the investigation of the estrogenic activity of synthetic chemicals (Fertuck et al 2001) and extracts of air particulates (Clemons et al 1998). The methodology has also allowed for incorporation into the screening evaluation of species-specific endocrine responses by application to ERs from varying organisms (Matthews et al 2002), although similar cross-species comparisons have not utilized this approach exclusively (Sumida et al 2003).…”

Section: Transactivation Assaysmentioning

confidence: 99%

In Vitro Models in Endocrine Disruptor Screening

Charles¹

2004

ILAR Journal

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The public and scientific concern that chemicals present in the human diet and the environment and their ability to disrupt the normal hormonal milieu in humans and wildlife have become a high-profile international issue. In 1998, the Endocrine Disruptor Screening and Testing Advisory Committee (EDSTAC) convened by the Environmental Protection Agency (EPA) recommended a tiered testing approach for the evaluation of estrogen, androgen, and thyroid-related effects of some 87,000 commercial chemicals and environmental contaminants. The function of this committee concluded with its final report, and the further implementation of the recommended testing strategy has now been carried forward with the assistance of the Endocrine Disruptor Methods Validation Subcommittee. The function of this body is to provide advice to the EPA on scientific and technical issues related specifically to the conduct of studies required for the validation of assays proposed by the EDSTAC as part of the tiered screening program. The EDSTAC recommended and alternative screening batteries encompass four in vitro mammalian assays. The current methodologies and validation status of the proposed in vitro EDSTAC assays are discussed and consist of estrogen/androgen receptor binding, estrogen/androgen gene transactivation, and minced testis, and one alternate (placental aromatase) in vitro screening assay.

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Limited species differences in estrogen receptor alpha-medicated reporter gene transactivation by xenoestrogens

Cited by 23 publications

References 29 publications

Molecular cloning of estrogen receptors from fathead minnow (Pimephales promelas) and bluegill (Lepomis macrochirus) fish: Limited piscine variation in estrogen receptor–mediated reporter gene transactivation by xenoestrogens

Molecular cloning of estrogen receptors from fathead minnow (Pimephales promelas) and bluegill (Lepomis macrochirus) fish: Limited piscine variation in estrogen receptor–mediated reporter gene transactivation by xenoestrogens

Comparison of chemical binding to recombinant fathead minnow and human estrogen receptors alpha in whole cell and cell-free binding assays

In Vitro Models in Endocrine Disruptor Screening

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