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17α‐ethynylestradiol‐induced vitellogenin gene transcription quantified in livers of adult males, larvae, and gills of fathead minnows ( Pimephales promelas )
Author(s) -
Lattier David L.,
Reddy Tirumuru V.,
Gordon Denise A.,
Lazorchak James M.,
Smith Mark E.,
Williams Daniel E.,
Wiechman Barry,
Flick Robert W.,
Miracle Ann L.,
Toth Gregory P.
Publication year - 2002
Publication title -
environmental toxicology and chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.1
H-Index - 171
eISSN - 1552-8618
pISSN - 0730-7268
DOI - 10.1002/etc.5620211118
Subject(s) - vitellogenin , pimephales promelas , minnow , biology , vitellogenins , gene expression , transcription (linguistics) , microbiology and biotechnology , gene , genetics , embryo , oocyte , fishery , vitellogenesis , fish <actinopterygii> , philosophy , linguistics
We have applied a method for quantifying relative levels of messenger RNA (mRNA) transcription to assess chemically induced gene expression in fathead minnows ( Pimephales promelas ). Synthetic oligonucleotides designed for the fathead minnow vitellogenin gene transcription product were used in a reverse transcription polymerase chain reaction (RT‐PCR) protocol. This sensitive and rapid strategy detected vitellogenin gene transcription in livers of male fathead minnows exposed to concentrations as low as 2 ng/L of the endocrine‐disrupting compound 17‐α‐ethynylestradiol for 24 h. Surprisingly, vitellogenin transcription products also were detected in gill tissue and in 48‐h‐old posthatch fathead minnow larvae. Relative levels of vitellogenin gene induction among individuals were quantified in a single‐step reaction (PCR multiplex) with 18S rRNA universal primers and Competimerst concurrently with fathead minnow vitellogenin oligonucleotides. This quantitative approach will markedly enhance detection of the first cellular event of estrogenic exposure to aquatic ecosystems in both field and laboratory systems. Use of the model provides sensitivity of detection at a concentration below those that cause mortality or visible signs of stress in fish or other aquatic organisms. The model may also provide an in vivo screening method for estrogenlike endocrine‐disrupting compounds.