Cloning, expression and functional characterization of carp, Cyprinus carpio, estrogen receptors and their differential activations by estrogens
Identifieur interne : 001632 ( Istex/Corpus ); précédent : 001631; suivant : 001633Cloning, expression and functional characterization of carp, Cyprinus carpio, estrogen receptors and their differential activations by estrogens
Auteurs : Yoshinao Katsu ; Anke Lange ; Shinichi Miyagawa ; Hiroshi Urushitani ; Norishisa Tatarazako ; Yukio Kawashima ; Charles R. Tyler ; Taisen IguchiSource :
- Journal of Applied Toxicology [ 0260-437X ] ; 2013-01.
Abstract
Sex‐steroid hormones are essential for normal reproductive activity in both sexes. Estrogens are necessary for ovarian differentiation during a critical developmental stage in vertebrates and promote the growth and differentiation of the female reproductive system. Importantly, environmental estrogens can influence the reproductive system and have been shown to disrupt gametogenesis in males. To understand the molecular mechanisms of estrogen actions and to evaluate estrogen receptor ligand interactions in the carp, Cyprinus carpio, a species used widely for both field‐ and laboratory‐based studies, we cloned all three carp estrogen receptors (ER; ERα, ERβ1 and ERβ2) and applied an estrogen‐responsive (ERE)‐luciferase reporter assay system to characterize the interactions of these receptors with steroidal and synthetic estrogens. DNA fragments encoding all three ERs in carp, ERα, ERβ1 and ERβ2, were obtained from the ovary using degenerate primer sets and PCR techniques, and full‐length carp ER (cER) cDNAs were then obtained using RACE (rapid amplification of the cDNA end) techniques. Amino acid sequences of cERs showed overall homology of 46% (α vs β1), 49% (α vs β2) and 53% (β1 vs β2). In the transient transfection ERE‐luciferase reporter assay system (using mammalian cells) the cER proteins displayed estrogen‐dependent activation of transcription and cERβ2 showed a higher sensitivity to the natural steroid oestrogen, 17β‐estradiol, than cERα. The assay system developed is a powerful assay for toxicology and provides a tool for future studies examining the receptor–environmental chemical interactions and estrogen‐disrupting mechanisms in carp. The data presented also expand our knowledge of estrogen receptor evolution. Copyright © 2011 John Wiley & Sons, Ltd.
Url:
DOI: 10.1002/jat.1707
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Ltd, 7‐5‐25 Nishi‐Shinjyuku, Tokyo, 160‐0023, Shinjyuku‐ku, Japan</mods:affiliation></affiliation></author><author><name sortKey="Tyler, Charles R" sort="Tyler, Charles R" uniqKey="Tyler C" first="Charles R." last="Tyler">Charles R. Tyler</name><affiliation><mods:affiliation>School of Biosciences, College of Life and Environmental Sciences, University of Exeter, EX4 4PS, Exeter, UK</mods:affiliation></affiliation></author><author><name sortKey="Iguchi, Taisen" sort="Iguchi, Taisen" uniqKey="Iguchi T" first="Taisen" last="Iguchi">Taisen Iguchi</name><affiliation><mods:affiliation>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5‐1 Higashiyama, Myodaiji, 444‐8787, Okazaki, Japan</mods:affiliation></affiliation><affiliation><mods:affiliation>Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies, 444‐8787, Okazaki, Japan</mods:affiliation></affiliation><affiliation><mods:affiliation>Taisen Iguchi, Okazaki Institutes for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5–1 Higashiyama, Myodaiji, Okazaki 444–8787, JapanE‐mail:</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: taisen@nibb.ac.jp</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Applied Toxicology</title><title level="j" type="abbrev">J. Appl. Toxicol.</title><idno type="ISSN">0260-437X</idno><idno type="eISSN">1099-1263</idno><imprint><publisher>John Wiley & Sons, Ltd</publisher><pubPlace>Chichester, UK</pubPlace><date type="published" when="2013-01">2013-01</date><biblScope unit="volume">33</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="41">41</biblScope><biblScope unit="page" to="49">49</biblScope></imprint><idno type="ISSN">0260-437X</idno></series><idno type="istex">43472B90A3A860CB58799672D48E4A751A64B667</idno><idno type="DOI">10.1002/jat.1707</idno><idno type="ArticleID">JAT1707</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0260-437X</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract">Sex‐steroid hormones are essential for normal reproductive activity in both sexes. Estrogens are necessary for ovarian differentiation during a critical developmental stage in vertebrates and promote the growth and differentiation of the female reproductive system. Importantly, environmental estrogens can influence the reproductive system and have been shown to disrupt gametogenesis in males. To understand the molecular mechanisms of estrogen actions and to evaluate estrogen receptor ligand interactions in the carp, Cyprinus carpio, a species used widely for both field‐ and laboratory‐based studies, we cloned all three carp estrogen receptors (ER; ERα, ERβ1 and ERβ2) and applied an estrogen‐responsive (ERE)‐luciferase reporter assay system to characterize the interactions of these receptors with steroidal and synthetic estrogens. DNA fragments encoding all three ERs in carp, ERα, ERβ1 and ERβ2, were obtained from the ovary using degenerate primer sets and PCR techniques, and full‐length carp ER (cER) cDNAs were then obtained using RACE (rapid amplification of the cDNA end) techniques. Amino acid sequences of cERs showed overall homology of 46% (α vs β1), 49% (α vs β2) and 53% (β1 vs β2). In the transient transfection ERE‐luciferase reporter assay system (using mammalian cells) the cER proteins displayed estrogen‐dependent activation of transcription and cERβ2 showed a higher sensitivity to the natural steroid oestrogen, 17β‐estradiol, than cERα. The assay system developed is a powerful assay for toxicology and provides a tool for future studies examining the receptor–environmental chemical interactions and estrogen‐disrupting mechanisms in carp. The data presented also expand our knowledge of estrogen receptor evolution. Copyright © 2011 John Wiley & Sons, Ltd.</div></front></TEI><istex><corpusName>wiley</corpusName><author><json:item><name>Yoshinao Katsu</name><affiliations><json:string>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5‐1 Higashiyama, Myodaiji, 444‐8787, Okazaki, Japan</json:string><json:string>Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies, 444‐8787, Okazaki, Japan</json:string><json:string>Department of Biological Sciences, Hokkaido University, 060‐0810, Sapporo, Japan</json:string></affiliations></json:item><json:item><name>Anke Lange</name><affiliations><json:string>School of Biosciences, College of Life and Environmental Sciences, University of Exeter, EX4 4PS, Exeter, UK</json:string></affiliations></json:item><json:item><name>Shinichi Miyagawa</name><affiliations><json:string>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5‐1 Higashiyama, Myodaiji, 444‐8787, Okazaki, Japan</json:string><json:string>Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies, 444‐8787, Okazaki, Japan</json:string></affiliations></json:item><json:item><name>Hiroshi Urushitani</name><affiliations><json:string>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5‐1 Higashiyama, Myodaiji, 444‐8787, Okazaki, Japan</json:string><json:string>National Institute for Environmental Studies, 16‐2 Onogawa, Ibaraki, 305‐8506, Tsukuba, Japan</json:string></affiliations></json:item><json:item><name>Norishisa Tatarazako</name><affiliations><json:string>National Institute for Environmental Studies, 16‐2 Onogawa, Ibaraki, 305‐8506, Tsukuba, Japan</json:string></affiliations></json:item><json:item><name>Yukio Kawashima</name><affiliations><json:string>Japan NUS Co. Ltd, 7‐5‐25 Nishi‐Shinjyuku, Tokyo, 160‐0023, Shinjyuku‐ku, Japan</json:string></affiliations></json:item><json:item><name>Charles R. Tyler</name><affiliations><json:string>School of Biosciences, College of Life and Environmental Sciences, University of Exeter, EX4 4PS, Exeter, UK</json:string></affiliations></json:item><json:item><name>Taisen Iguchi</name><affiliations><json:string>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5‐1 Higashiyama, Myodaiji, 444‐8787, Okazaki, Japan</json:string><json:string>Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies, 444‐8787, Okazaki, Japan</json:string><json:string>Taisen Iguchi, Okazaki Institutes for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5–1 Higashiyama, Myodaiji, Okazaki 444–8787, JapanE‐mail:</json:string><json:string>E-mail: taisen@nibb.ac.jp</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>carp</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>estrogen receptors</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>transactivation assay</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>estrogens</value></json:item></subject><articleId><json:string>JAT1707</json:string></articleId><language><json:string>eng</json:string></language><originalGenre><json:string>article</json:string></originalGenre><qualityIndicators><score>8.5</score><pdfVersion>1.4</pdfVersion><pdfPageSize>595.276 x 790.866 pts</pdfPageSize><refBibsNative>true</refBibsNative><abstractCharCount>1791</abstractCharCount><pdfWordCount>6998</pdfWordCount><pdfCharCount>45368</pdfCharCount><pdfPageCount>9</pdfPageCount><abstractWordCount>257</abstractWordCount></qualityIndicators><title>Cloning, 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their differential activations by estrogens</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>John Wiley & Sons, Ltd</publisher><pubPlace>Chichester, UK</pubPlace><availability><p>Copyright © 2012 John Wiley & Sons, Ltd.Copyright © 2011 John Wiley & Sons, Ltd.</p></availability><date>2011-06-01</date></publicationStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a" type="main" xml:lang="en">Cloning, expression and functional characterization of carp, Cyprinus carpio, estrogen receptors and their differential activations by estrogens</title><author xml:id="author-1"><persName><forename type="first">Yoshinao</forename><surname>Katsu</surname></persName><affiliation>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5‐1 Higashiyama, Myodaiji, 444‐8787, Okazaki, Japan</affiliation><affiliation>Department of Basic Biology, School of Life Science, Graduate University for 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type="first">Hiroshi</forename><surname>Urushitani</surname></persName><affiliation>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5‐1 Higashiyama, Myodaiji, 444‐8787, Okazaki, Japan</affiliation><affiliation>National Institute for Environmental Studies, 16‐2 Onogawa, Ibaraki, 305‐8506, Tsukuba, Japan</affiliation></author><author xml:id="author-5"><persName><forename type="first">Norishisa</forename><surname>Tatarazako</surname></persName><affiliation>National Institute for Environmental Studies, 16‐2 Onogawa, Ibaraki, 305‐8506, Tsukuba, Japan</affiliation></author><author xml:id="author-6"><persName><forename type="first">Yukio</forename><surname>Kawashima</surname></persName><affiliation>Japan NUS Co. Ltd, 7‐5‐25 Nishi‐Shinjyuku, Tokyo, 160‐0023, Shinjyuku‐ku, Japan</affiliation></author><author xml:id="author-7"><persName><forename type="first">Charles R.</forename><surname>Tyler</surname></persName><affiliation>School of Biosciences, College of Life and Environmental Sciences, University of Exeter, EX4 4PS, Exeter, UK</affiliation></author><author xml:id="author-8"><persName><forename type="first">Taisen</forename><surname>Iguchi</surname></persName><email>taisen@nibb.ac.jp</email><affiliation>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5‐1 Higashiyama, Myodaiji, 444‐8787, Okazaki, Japan</affiliation><affiliation>Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies, 444‐8787, Okazaki, Japan</affiliation><affiliation>Taisen Iguchi, Okazaki Institutes for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5–1 Higashiyama, Myodaiji, Okazaki 444–8787, JapanE‐mail:</affiliation></author></analytic><monogr><title level="j">Journal of Applied Toxicology</title><title level="j" type="abbrev">J. Appl. Toxicol.</title><idno type="pISSN">0260-437X</idno><idno type="eISSN">1099-1263</idno><idno type="DOI">10.1002/(ISSN)1099-1263</idno><imprint><publisher>John Wiley & Sons, Ltd</publisher><pubPlace>Chichester, UK</pubPlace><date type="published" when="2013-01"></date><biblScope unit="volume">33</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="41">41</biblScope><biblScope unit="page" to="49">49</biblScope></imprint></monogr><idno type="istex">43472B90A3A860CB58799672D48E4A751A64B667</idno><idno type="DOI">10.1002/jat.1707</idno><idno type="ArticleID">JAT1707</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>2011-06-01</date></creation><langUsage><language ident="en">en</language></langUsage><abstract><p>Sex‐steroid hormones are essential for normal reproductive activity in both sexes. Estrogens are necessary for ovarian differentiation during a critical developmental stage in vertebrates and promote the growth and differentiation of the female reproductive system. Importantly, environmental estrogens can influence the reproductive system and have been shown to disrupt gametogenesis in males. To understand the molecular mechanisms of estrogen actions and to evaluate estrogen receptor ligand interactions in the carp, Cyprinus carpio, a species used widely for both field‐ and laboratory‐based studies, we cloned all three carp estrogen receptors (ER; ERα, ERβ1 and ERβ2) and applied an estrogen‐responsive (ERE)‐luciferase reporter assay system to characterize the interactions of these receptors with steroidal and synthetic estrogens. DNA fragments encoding all three ERs in carp, ERα, ERβ1 and ERβ2, were obtained from the ovary using degenerate primer sets and PCR techniques, and full‐length carp ER (cER) cDNAs were then obtained using RACE (rapid amplification of the cDNA end) techniques. Amino acid sequences of cERs showed overall homology of 46% (α vs β1), 49% (α vs β2) and 53% (β1 vs β2). In the transient transfection ERE‐luciferase reporter assay system (using mammalian cells) the cER proteins displayed estrogen‐dependent activation of transcription and cERβ2 showed a higher sensitivity to the natural steroid oestrogen, 17β‐estradiol, than cERα. The assay system developed is a powerful assay for toxicology and provides a tool for future studies examining the receptor–environmental chemical interactions and estrogen‐disrupting mechanisms in carp. The data presented also expand our knowledge of estrogen receptor evolution. Copyright © 2011 John Wiley & Sons, Ltd.</p></abstract><abstract style="short"><p>Environmental estrogens can influence the reproductive system and have been shown to disrupt gametogenesis in males. We cloned all three carp Cyprinus carpio, estrogen receptors [ER; ERa, ERb1 and ERb2] and applied a transient transfection ERE‐luciferase reporter assay system using mammalian cells. cERb2 showed a higher sensitivity to the natural steroid oestrogen, 17b‐estradiol, than cERa. This is a powerful assay for toxicology, and provides a tool for future studies examining the receptor‐environmental chemical interactions and estrogen disrupting mechanisms in carp.</p></abstract><textClass><keywords scheme="keyword"><list><head>keywords</head><item><term>carp</term></item><item><term>estrogen receptors</term></item><item><term>transactivation assay</term></item><item><term>estrogens</term></item></list></keywords></textClass><textClass><keywords scheme="Journal Subject"><list><head>article-category</head><item><term>Research Article</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="2011-03-17">Received</change><change when="2011-05-07">Registration</change><change when="2011-06-01">Created</change><change when="2013-01">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/document/43472B90A3A860CB58799672D48E4A751A64B667/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Wiley, elements deleted: body"><istex:xmlDeclaration>version="1.0" encoding="UTF-8" standalone="yes"</istex:xmlDeclaration><istex:document><component type="serialArticle" version="2.0" xml:lang="en" xml:id="jat1707"><header><publicationMeta level="product"><publisherInfo><publisherName>John Wiley & Sons, Ltd</publisherName><publisherLoc>Chichester, UK</publisherLoc></publisherInfo><doi>10.1002/(ISSN)1099-1263</doi><issn type="print">0260-437X</issn><issn type="electronic">1099-1263</issn><idGroup><id type="product" value="JAT"></id></idGroup><titleGroup><title type="main" sort="JOURNAL OF APPLIED TOXICOLOGY">Journal of Applied Toxicology</title><title type="short">J. Appl. Toxicol.</title></titleGroup></publicationMeta><publicationMeta level="part" position="10"><doi>10.1002/jat.v33.1</doi><copyright ownership="publisher">Copyright © 2012 John Wiley & Sons, Ltd.</copyright><numberingGroup><numbering type="journalVolume" number="33">33</numbering><numbering type="journalIssue">1</numbering></numberingGroup><coverDate startDate="2013-01">January 2013</coverDate></publicationMeta><publicationMeta level="unit" position="60" type="article" status="forIssue"><doi>10.1002/jat.1707</doi><idGroup><id type="unit" value="JAT1707"></id></idGroup><countGroup><count type="pageTotal" number="9"></count></countGroup><titleGroup><title type="articleCategory">Research Article</title><title type="tocHeading1">Research Articles</title></titleGroup><copyright ownership="publisher">Copyright © 2011 John Wiley & Sons, Ltd.</copyright><eventGroup><event type="manuscriptReceived" date="2011-03-17"></event><event type="manuscriptRevised" date="2011-05-07"></event><event type="manuscriptAccepted" date="2011-05-07"></event><event type="xmlCreated" agent="SPi Global" date="2011-06-01"></event><event type="publishedOnlineEarlyUnpaginated" date="2011-07-01"></event><event type="firstOnline" date="2011-07-01"></event><event type="publishedOnlineFinalForm" date="2012-11-29"></event><event type="xmlConverted" agent="Converter:WILEY_ML3G_TO_WILEY_ML3GV2 version:3.8.8" date="2014-01-28"></event><event type="xmlConverted" agent="Converter:WML3G_To_WML3G version:4.6.4 mode:FullText" date="2015-10-04"></event></eventGroup><numberingGroup><numbering type="pageFirst">41</numbering><numbering type="pageLast">49</numbering></numberingGroup><correspondenceTo><lineatedText><line>Taisen Iguchi, Okazaki Institutes for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5–1 Higashiyama, Myodaiji, Okazaki 444–8787, Japan</line><line>E‐mail: <email>taisen@nibb.ac.jp</email></line></lineatedText></correspondenceTo><linkGroup><link type="toTypesetVersion" href="file:JAT.JAT1707.pdf"></link></linkGroup></publicationMeta><contentMeta><titleGroup><title type="shortAuthors">Y. Katsu <i>et al.</i></title><title type="main">Cloning, expression and functional characterization of carp, <i>Cyprinus carpio</i>, estrogen receptors and their differential activations by estrogens</title><title type="short">Functional characterization of carp estrogen receptors</title></titleGroup><creators><creator creatorRole="author" xml:id="jat1707-cr-0001" affiliationRef="#jat1707-aff-0001 #jat1707-aff-0002 #jat1707-aff-0003"><personName><givenNames>Yoshinao</givenNames><familyName>Katsu</familyName></personName></creator><creator creatorRole="author" xml:id="jat1707-cr-0002" affiliationRef="#jat1707-aff-0004"><personName><givenNames>Anke</givenNames><familyName>Lange</familyName></personName></creator><creator creatorRole="author" xml:id="jat1707-cr-0003" affiliationRef="#jat1707-aff-0001 #jat1707-aff-0002"><personName><givenNames>Shinichi</givenNames><familyName>Miyagawa</familyName></personName></creator><creator creatorRole="author" xml:id="jat1707-cr-0004" affiliationRef="#jat1707-aff-0001 #jat1707-aff-0005"><personName><givenNames>Hiroshi</givenNames><familyName>Urushitani</familyName></personName></creator><creator creatorRole="author" xml:id="jat1707-cr-0005" affiliationRef="#jat1707-aff-0005"><personName><givenNames>Norishisa</givenNames><familyName>Tatarazako</familyName></personName></creator><creator creatorRole="author" xml:id="jat1707-cr-0006" affiliationRef="#jat1707-aff-0006"><personName><givenNames>Yukio</givenNames><familyName>Kawashima</familyName></personName></creator><creator creatorRole="author" xml:id="jat1707-cr-0007" affiliationRef="#jat1707-aff-0004"><personName><givenNames>Charles R.</givenNames><familyName>Tyler</familyName></personName></creator><creator creatorRole="author" xml:id="jat1707-cr-0008" affiliationRef="#jat1707-aff-0001 #jat1707-aff-0002" corresponding="yes"><personName><givenNames>Taisen</givenNames><familyName>Iguchi</familyName></personName><contactDetails><email>taisen@nibb.ac.jp</email></contactDetails></creator></creators><affiliationGroup><affiliation xml:id="jat1707-aff-0001" countryCode="JP" type="organization"><orgDiv>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology</orgDiv><orgName>National Institutes of Natural Sciences</orgName><address><street>5‐1 Higashiyama, Myodaiji</street><city>Okazaki</city><postCode>444‐8787</postCode><country>Japan</country></address></affiliation><affiliation xml:id="jat1707-aff-0002" countryCode="JP" type="organization"><orgDiv>Department of Basic Biology, School of Life Science</orgDiv><orgName>Graduate University for Advanced Studies</orgName><address><city>Okazaki</city><postCode>444‐8787</postCode><country>Japan</country></address></affiliation><affiliation xml:id="jat1707-aff-0003" countryCode="JP" type="organization"><orgDiv>Department of Biological Sciences</orgDiv><orgName>Hokkaido University</orgName><address><city>Sapporo</city><postCode>060‐0810</postCode><country>Japan</country></address></affiliation><affiliation xml:id="jat1707-aff-0004" countryCode="GB" type="organization"><orgDiv>School of Biosciences, College of Life and Environmental Sciences</orgDiv><orgName>University of Exeter</orgName><address><city>Exeter</city><postCode>EX4 4PS</postCode><country>UK</country></address></affiliation><affiliation xml:id="jat1707-aff-0005" countryCode="JP" type="organization"><orgName>National Institute for Environmental Studies</orgName><address><street>16‐2 Onogawa</street><city>Tsukuba</city><countryPart>Ibaraki</countryPart><postCode>305‐8506</postCode><country>Japan</country></address></affiliation><affiliation xml:id="jat1707-aff-0006" countryCode="JP" type="organization"><orgName>Japan NUS Co. Ltd, 7‐5‐25 Nishi‐Shinjyuku</orgName><address><city>Shinjyuku‐ku</city><countryPart>Tokyo</countryPart><postCode>160‐0023</postCode><country>Japan</country></address></affiliation></affiliationGroup><keywordGroup type="author"><keyword xml:id="jat1707-kwd-0001">carp</keyword><keyword xml:id="jat1707-kwd-0002">estrogen receptors</keyword><keyword xml:id="jat1707-kwd-0003">transactivation assay</keyword><keyword xml:id="jat1707-kwd-0004">estrogens</keyword></keywordGroup><abstractGroup><abstract type="main"><title type="main">ABSTRACT</title><p>Sex‐steroid hormones are essential for normal reproductive activity in both sexes. Estrogens are necessary for ovarian differentiation during a critical developmental stage in vertebrates and promote the growth and differentiation of the female reproductive system. Importantly, environmental estrogens can influence the reproductive system and have been shown to disrupt gametogenesis in males. To understand the molecular mechanisms of estrogen actions and to evaluate estrogen receptor ligand interactions in the carp, <i>Cyprinus carpio</i>, a species used widely for both field‐ and laboratory‐based studies, we cloned all three carp estrogen receptors (ER; ERα, ERβ1 and ERβ2) and applied an estrogen‐responsive (ERE)‐luciferase reporter assay system to characterize the interactions of these receptors with steroidal and synthetic estrogens. DNA fragments encoding all three ERs in carp, ERα, ERβ1 and ERβ2, were obtained from the ovary using degenerate primer sets and PCR techniques, and full‐length carp ER (cER) cDNAs were then obtained using RACE (rapid amplification of the cDNA end) techniques. Amino acid sequences of cERs showed overall homology of 46% (α vs β1), 49% (α vs β2) and 53% (β1 vs β2). In the transient transfection ERE‐luciferase reporter assay system (using mammalian cells) the cER proteins displayed estrogen‐dependent activation of transcription and cERβ2 showed a higher sensitivity to the natural steroid oestrogen, 17β‐estradiol, than cERα. The assay system developed is a powerful assay for toxicology and provides a tool for future studies examining the receptor–environmental chemical interactions and estrogen‐disrupting mechanisms in carp. The data presented also expand our knowledge of estrogen receptor evolution. Copyright © 2011 John Wiley & Sons, Ltd.</p></abstract><abstract type="short" xml:id="jat1707-abs-0001"><p>Environmental estrogens can influence the reproductive system and have been shown to disrupt gametogenesis in males. We cloned all three carp <i>Cyprinus carpio</i>, estrogen receptors [ER; ERa, ERb1 and ERb2] and applied a transient transfection ERE‐luciferase reporter assay system using mammalian cells. cERb2 showed a higher sensitivity to the natural steroid oestrogen, 17b‐estradiol, than cERa. This is a powerful assay for toxicology, and provides a tool for future studies examining the receptor‐environmental chemical interactions and estrogen disrupting mechanisms in carp.</p></abstract></abstractGroup></contentMeta></header></component></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Cloning, expression and functional characterization of carp, Cyprinus carpio, estrogen receptors and their differential activations by estrogens</title></titleInfo><titleInfo type="abbreviated" lang="en"><title>Functional characterization of carp estrogen receptors</title></titleInfo><titleInfo type="alternative" contentType="CDATA" lang="en"><title>Cloning, expression and functional characterization of carp, Cyprinus carpio, estrogen receptors and their differential activations by estrogens</title></titleInfo><name type="personal"><namePart type="given">Yoshinao</namePart><namePart type="family">Katsu</namePart><affiliation>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5‐1 Higashiyama, Myodaiji, 444‐8787, Okazaki, Japan</affiliation><affiliation>Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies, 444‐8787, Okazaki, Japan</affiliation><affiliation>Department of Biological Sciences, Hokkaido University, 060‐0810, Sapporo, Japan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Anke</namePart><namePart type="family">Lange</namePart><affiliation>School of Biosciences, College of Life and Environmental Sciences, University of Exeter, EX4 4PS, Exeter, UK</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Shinichi</namePart><namePart type="family">Miyagawa</namePart><affiliation>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5‐1 Higashiyama, Myodaiji, 444‐8787, Okazaki, Japan</affiliation><affiliation>Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies, 444‐8787, Okazaki, Japan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Hiroshi</namePart><namePart type="family">Urushitani</namePart><affiliation>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5‐1 Higashiyama, Myodaiji, 444‐8787, Okazaki, Japan</affiliation><affiliation>National Institute for Environmental Studies, 16‐2 Onogawa, Ibaraki, 305‐8506, Tsukuba, Japan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Norishisa</namePart><namePart type="family">Tatarazako</namePart><affiliation>National Institute for Environmental Studies, 16‐2 Onogawa, Ibaraki, 305‐8506, Tsukuba, Japan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Yukio</namePart><namePart type="family">Kawashima</namePart><affiliation>Japan NUS Co. Ltd, 7‐5‐25 Nishi‐Shinjyuku, Tokyo, 160‐0023, Shinjyuku‐ku, Japan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Charles R.</namePart><namePart type="family">Tyler</namePart><affiliation>School of Biosciences, College of Life and Environmental Sciences, University of Exeter, EX4 4PS, Exeter, UK</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Taisen</namePart><namePart type="family">Iguchi</namePart><affiliation>Okazaki Institute for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5‐1 Higashiyama, Myodaiji, 444‐8787, Okazaki, Japan</affiliation><affiliation>Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies, 444‐8787, Okazaki, Japan</affiliation><affiliation>Taisen Iguchi, Okazaki Institutes for Integrative Bioscience, National Institute for Basic Biology, National Institutes of Natural Sciences, 5–1 Higashiyama, Myodaiji, Okazaki 444–8787, JapanE‐mail:</affiliation><affiliation>E-mail: taisen@nibb.ac.jp</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="article" displayLabel="article"></genre><originInfo><publisher>John Wiley & Sons, Ltd</publisher><place><placeTerm type="text">Chichester, UK</placeTerm></place><dateIssued encoding="w3cdtf">2013-01</dateIssued><dateCreated encoding="w3cdtf">2011-06-01</dateCreated><dateCaptured encoding="w3cdtf">2011-03-17</dateCaptured><dateValid encoding="w3cdtf">2011-05-07</dateValid><copyrightDate encoding="w3cdtf">2013</copyrightDate></originInfo><language><languageTerm type="code" authority="rfc3066">en</languageTerm><languageTerm type="code" authority="iso639-2b">eng</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract>Sex‐steroid hormones are essential for normal reproductive activity in both sexes. Estrogens are necessary for ovarian differentiation during a critical developmental stage in vertebrates and promote the growth and differentiation of the female reproductive system. Importantly, environmental estrogens can influence the reproductive system and have been shown to disrupt gametogenesis in males. To understand the molecular mechanisms of estrogen actions and to evaluate estrogen receptor ligand interactions in the carp, Cyprinus carpio, a species used widely for both field‐ and laboratory‐based studies, we cloned all three carp estrogen receptors (ER; ERα, ERβ1 and ERβ2) and applied an estrogen‐responsive (ERE)‐luciferase reporter assay system to characterize the interactions of these receptors with steroidal and synthetic estrogens. DNA fragments encoding all three ERs in carp, ERα, ERβ1 and ERβ2, were obtained from the ovary using degenerate primer sets and PCR techniques, and full‐length carp ER (cER) cDNAs were then obtained using RACE (rapid amplification of the cDNA end) techniques. Amino acid sequences of cERs showed overall homology of 46% (α vs β1), 49% (α vs β2) and 53% (β1 vs β2). In the transient transfection ERE‐luciferase reporter assay system (using mammalian cells) the cER proteins displayed estrogen‐dependent activation of transcription and cERβ2 showed a higher sensitivity to the natural steroid oestrogen, 17β‐estradiol, than cERα. The assay system developed is a powerful assay for toxicology and provides a tool for future studies examining the receptor–environmental chemical interactions and estrogen‐disrupting mechanisms in carp. The data presented also expand our knowledge of estrogen receptor evolution. Copyright © 2011 John Wiley & Sons, Ltd.</abstract><abstract type="short">Environmental estrogens can influence the reproductive system and have been shown to disrupt gametogenesis in males. We cloned all three carp Cyprinus carpio, estrogen receptors [ER; ERa, ERb1 and ERb2] and applied a transient transfection ERE‐luciferase reporter assay system using mammalian cells. cERb2 showed a higher sensitivity to the natural steroid oestrogen, 17b‐estradiol, than cERa. This is a powerful assay for toxicology, and provides a tool for future studies examining the receptor‐environmental chemical interactions and estrogen disrupting mechanisms in carp.</abstract><subject><genre>keywords</genre><topic>carp</topic><topic>estrogen receptors</topic><topic>transactivation assay</topic><topic>estrogens</topic></subject><relatedItem type="host"><titleInfo><title>Journal of Applied Toxicology</title></titleInfo><titleInfo type="abbreviated"><title>J. Appl. Toxicol.</title></titleInfo><genre type="journal">journal</genre><subject><genre>article-category</genre><topic>Research Article</topic></subject><identifier type="ISSN">0260-437X</identifier><identifier type="eISSN">1099-1263</identifier><identifier type="DOI">10.1002/(ISSN)1099-1263</identifier><identifier type="PublisherID">JAT</identifier><part><date>2013</date><detail type="volume"><caption>vol.</caption><number>33</number></detail><detail type="issue"><caption>no.</caption><number>1</number></detail><extent unit="pages"><start>41</start><end>49</end><total>9</total></extent></part></relatedItem><identifier type="istex">43472B90A3A860CB58799672D48E4A751A64B667</identifier><identifier type="DOI">10.1002/jat.1707</identifier><identifier type="ArticleID">JAT1707</identifier><accessCondition type="use and reproduction" contentType="copyright">Copyright © 2012 John Wiley & Sons, Ltd.Copyright © 2011 John Wiley & Sons, Ltd.</accessCondition><recordInfo><recordContentSource>WILEY</recordContentSource><recordOrigin>John Wiley & Sons, Ltd</recordOrigin></recordInfo></mods></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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