Fathead minnow vitellogenin: Complementary DNA sequence and messenger RNA and protein expression after 17β‐estradiol treatment
Identifieur interne : 001283 ( Istex/Corpus ); précédent : 001282; suivant : 001284Fathead minnow vitellogenin: Complementary DNA sequence and messenger RNA and protein expression after 17β‐estradiol treatment
Auteurs : Joseph J. Korte ; Michael D. Kahl ; Kathleen M. Jensen ; Mumtaz S. Pasha ; Louise G. Parks ; Gerald A. Leblanc ; Gerald T. AnkleySource :
- Environmental Toxicology and Chemistry [ 0730-7268 ] ; 2000-04.
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- KwdEn :
Abstract
Induction of vitellogenin (VTG) in oviparous animals has been proposed as a sensitive indicator of environmental contaminants that activate the estrogen receptor. In the present study, a sensitive ribonuclease protection assay (RPA) for VTG messenger RNA (mRNA) was developed for the fathead minnow (Pimephales promelas), a species proposed for routine endocrine‐disrupting chemical (EDC) screening. The utility of this method was compared with an enzyme‐linked immunosorbent assay (ELISA) specific for fathead minnow VTG protein. Assessment of the two methods included kinetic characterization of the plasma VTG protein and hepatic VTG mRNA levels in male fathead minnows following intraperitoneal injections of 17β‐estradiol (E2) at two dose levels (0.5, 5.0 mg/kg). Initial plasma E2 concentrations were elevated in a dose‐dependent manner but returned to normal levels within 2 d. Liver VTG mRNA was detected within 4 h, reached a maximum around 48 h, and returned to normal levels in about 6 d. Plasma VTG protein was detectable within 16 h of treatment, reached maximum levels at about 72 h, and remained near these maximum levels for at least 18 d. While the RPA was about 1,000 times more sensitive than the ELISA, the ELISA appears superior for routine screening tests. The ELISA method is relatively simple to perform and, because males lack a clearance mechanism for VTG, the protein remains at relatively high concentrations in the plasma for an extended period of time. As part of the development of the RPA, the complementary DNA (cDNA) sequence for fathead minnow VTG was determined and the deduced amino acid sequence compared with VTG sequences for other fish species.
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DOI: 10.1002/etc.5620190426
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<front><div type="abstract" xml:lang="en">Induction of vitellogenin (VTG) in oviparous animals has been proposed as a sensitive indicator of environmental contaminants that activate the estrogen receptor. In the present study, a sensitive ribonuclease protection assay (RPA) for VTG messenger RNA (mRNA) was developed for the fathead minnow (Pimephales promelas), a species proposed for routine endocrine‐disrupting chemical (EDC) screening. The utility of this method was compared with an enzyme‐linked immunosorbent assay (ELISA) specific for fathead minnow VTG protein. Assessment of the two methods included kinetic characterization of the plasma VTG protein and hepatic VTG mRNA levels in male fathead minnows following intraperitoneal injections of 17β‐estradiol (E2) at two dose levels (0.5, 5.0 mg/kg). Initial plasma E2 concentrations were elevated in a dose‐dependent manner but returned to normal levels within 2 d. Liver VTG mRNA was detected within 4 h, reached a maximum around 48 h, and returned to normal levels in about 6 d. Plasma VTG protein was detectable within 16 h of treatment, reached maximum levels at about 72 h, and remained near these maximum levels for at least 18 d. While the RPA was about 1,000 times more sensitive than the ELISA, the ELISA appears superior for routine screening tests. The ELISA method is relatively simple to perform and, because males lack a clearance mechanism for VTG, the protein remains at relatively high concentrations in the plasma for an extended period of time. As part of the development of the RPA, the complementary DNA (cDNA) sequence for fathead minnow VTG was determined and the deduced amino acid sequence compared with VTG sequences for other fish species.</div>
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<titleGroup><title type="articleCategory">Environmental Toxicology</title>
<title type="tocHeading1">Environmental Toxicology</title>
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<copyright ownership="thirdParty">Copyright © 2000 SETAC</copyright>
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<correspondenceTo>U.S. Environmental Protection Agency, Mid‐Continent Ecology Division, 6201 Congdon Boulevard, Duluth, Minnesota 55804</correspondenceTo>
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<titleGroup><title type="main" xml:lang="en">Fathead minnow vitellogenin: Complementary DNA sequence and messenger RNA and protein expression after 17β‐estradiol treatment<link href="#fn1"></link>
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<title type="short" xml:lang="en">Expression of fathead minnow vitellogenin mRNA and protein</title>
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<creators><creator xml:id="au1" creatorRole="author" affiliationRef="#af1" corresponding="yes"><personName><givenNames>Joseph J.</givenNames>
<familyName>Korte</familyName>
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<contactDetails><email>korte.joe@epamail.epa.gov</email>
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<creator xml:id="au2" creatorRole="author" affiliationRef="#af1"><personName><givenNames>Michael D.</givenNames>
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<creator xml:id="au3" creatorRole="author" affiliationRef="#af1"><personName><givenNames>Kathleen M.</givenNames>
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<creator xml:id="au4" creatorRole="author" affiliationRef="#af1"><personName><givenNames>Mumtaz S.</givenNames>
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<creator xml:id="au5" creatorRole="author" affiliationRef="#af2"><personName><givenNames>Louise G.</givenNames>
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<creator xml:id="au6" creatorRole="author" affiliationRef="#af2"><personName><givenNames>Gerald A.</givenNames>
<familyName>LeBlanc</familyName>
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<creator xml:id="au7" creatorRole="author" affiliationRef="#af1"><personName><givenNames>Gerald T.</givenNames>
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<affiliation xml:id="af2" countryCode="US" type="organization"><unparsedAffiliation>North Carolina State University Department of Toxicology, Box 7633, Raleigh, North Carolina 27695, USA</unparsedAffiliation>
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<keywordGroup xml:lang="en" type="author"><keyword xml:id="kwd1">Vitellogenin</keyword>
<keyword xml:id="kwd2"><i>Pimephales promelas</i>
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<keyword xml:id="kwd3">17β‐Estradiol</keyword>
<keyword xml:id="kwd4">mRNA</keyword>
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<abstractGroup><abstract type="main" xml:lang="en"><title type="main">Abstract</title>
<p>Induction of vitellogenin (VTG) in oviparous animals has been proposed as a sensitive indicator of environmental contaminants that activate the estrogen receptor. In the present study, a sensitive ribonuclease protection assay (RPA) for VTG messenger RNA (mRNA) was developed for the fathead minnow (<i>Pimephales promelas</i>
), a species proposed for routine endocrine‐disrupting chemical (EDC) screening. The utility of this method was compared with an enzyme‐linked immunosorbent assay (ELISA) specific for fathead minnow VTG protein. Assessment of the two methods included kinetic characterization of the plasma VTG protein and hepatic VTG mRNA levels in male fathead minnows following intraperitoneal injections of 17β‐estradiol (E2) at two dose levels (0.5, 5.0 mg/kg). Initial plasma E2 concentrations were elevated in a dose‐dependent manner but returned to normal levels within 2 d. Liver VTG mRNA was detected within 4 h, reached a maximum around 48 h, and returned to normal levels in about 6 d. Plasma VTG protein was detectable within 16 h of treatment, reached maximum levels at about 72 h, and remained near these maximum levels for at least 18 d. While the RPA was about 1,000 times more sensitive than the ELISA, the ELISA appears superior for routine screening tests. The ELISA method is relatively simple to perform and, because males lack a clearance mechanism for VTG, the protein remains at relatively high concentrations in the plasma for an extended period of time. As part of the development of the RPA, the complementary DNA (cDNA) sequence for fathead minnow VTG was determined and the deduced amino acid sequence compared with VTG sequences for other fish species.</p>
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<noteGroup><note xml:id="fn1"><p>This manuscript has been reviewed in accordance with official U.S. Environmental Protection Agency (U.S. EPA) procedures; however, the content does not reflect U.S. EPA policy. Mention of trade names does not imply endorsement by the U.S. EPA or the U.S. government.</p>
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<name type="personal"><namePart type="given">Joseph J.</namePart>
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<affiliation>U.S. Environmental Protection Agency, Mid‐Continent Ecology Division, 6201 Congdon Boulevard, Duluth, Minnesota 55804</affiliation>
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<name type="personal"><namePart type="given">Michael D.</namePart>
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<role><roleTerm type="text">author</roleTerm>
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<name type="personal"><namePart type="given">Kathleen M.</namePart>
<namePart type="family">Jensen</namePart>
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<role><roleTerm type="text">author</roleTerm>
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<name type="personal"><namePart type="given">Mumtaz S.</namePart>
<namePart type="family">Pasha</namePart>
<affiliation>U.S. Environmental Protection Agency, Mid‐Continent Ecology Division, 6201 Congdon Boulevard, Duluth, Minnesota 55804</affiliation>
<role><roleTerm type="text">author</roleTerm>
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<name type="personal"><namePart type="given">Louise G.</namePart>
<namePart type="family">Parks</namePart>
<affiliation>North Carolina State University Department of Toxicology, Box 7633, Raleigh, North Carolina 27695, USA</affiliation>
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<name type="personal"><namePart type="given">Gerald A.</namePart>
<namePart type="family">LeBlanc</namePart>
<affiliation>North Carolina State University Department of Toxicology, Box 7633, Raleigh, North Carolina 27695, USA</affiliation>
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<name type="personal"><namePart type="given">Gerald T.</namePart>
<namePart type="family">Ankley</namePart>
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<abstract lang="en">Induction of vitellogenin (VTG) in oviparous animals has been proposed as a sensitive indicator of environmental contaminants that activate the estrogen receptor. In the present study, a sensitive ribonuclease protection assay (RPA) for VTG messenger RNA (mRNA) was developed for the fathead minnow (Pimephales promelas), a species proposed for routine endocrine‐disrupting chemical (EDC) screening. The utility of this method was compared with an enzyme‐linked immunosorbent assay (ELISA) specific for fathead minnow VTG protein. Assessment of the two methods included kinetic characterization of the plasma VTG protein and hepatic VTG mRNA levels in male fathead minnows following intraperitoneal injections of 17β‐estradiol (E2) at two dose levels (0.5, 5.0 mg/kg). Initial plasma E2 concentrations were elevated in a dose‐dependent manner but returned to normal levels within 2 d. Liver VTG mRNA was detected within 4 h, reached a maximum around 48 h, and returned to normal levels in about 6 d. Plasma VTG protein was detectable within 16 h of treatment, reached maximum levels at about 72 h, and remained near these maximum levels for at least 18 d. While the RPA was about 1,000 times more sensitive than the ELISA, the ELISA appears superior for routine screening tests. The ELISA method is relatively simple to perform and, because males lack a clearance mechanism for VTG, the protein remains at relatively high concentrations in the plasma for an extended period of time. As part of the development of the RPA, the complementary DNA (cDNA) sequence for fathead minnow VTG was determined and the deduced amino acid sequence compared with VTG sequences for other fish species.</abstract>
<note type="content">*This manuscript has been reviewed in accordance with official U.S. Environmental Protection Agency (U.S. EPA) procedures; however, the content does not reflect U.S. EPA policy. Mention of trade names does not imply endorsement by the U.S. EPA or the U.S. government.</note>
<subject lang="en"><genre>keywords</genre>
<topic>Vitellogenin</topic>
<topic>Pimephales promelas</topic>
<topic>17β‐Estradiol</topic>
<topic>mRNA</topic>
<topic>cDNA</topic>
</subject>
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<subTitle>An International Journal</subTitle>
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<identifier type="ISSN">0730-7268</identifier>
<identifier type="eISSN">1552-8618</identifier>
<identifier type="DOI">10.1002/(ISSN)1552-8618</identifier>
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<part><date>2000</date>
<detail type="volume"><caption>vol.</caption>
<number>19</number>
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