Vitellogenin gene transcription: A relative quantitative exposure indicator of environmental estrogens

Lattier, David L.; Gordon, Denise A.; Burks, Douglas J.; Toth, Gregory P.

doi:10.1002/etc.5620200916

Cited by 39 publications

(11 citation statements)

References 45 publications

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“…The data shown here confirm the current trend in the use of the methods of advanced molecular biology to study the effects of environmental pollutants on exposed biota [19,20,26,27,28,29]. RNA-based methods have several advantages over protein-based methods.…”

Section: Discussionsupporting

confidence: 84%

“…Vg is synthesised in the liver under the control of 17β-estradiol (estradiol), secreted into the blood stream, and finally taken up by oocytes via receptor-mediated endocytosis. Vg synthesis normally occurs in sexually active females, although it can also occur in male and immature animals in response to exogenous estrogenic substances [16,17,18,19,20].…”

Section: Introductionmentioning

confidence: 99%

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Use of vitellogenin mRNA as a biomarker for endocrine disruption in feral and cultured fish

García-Reyero¹,

Raldúa

Quirós

et al. 2004

Analytical and Bioanalytical Chemistry

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The presence of the female-specific yolk protein precursor vitellogenin in blood and liver from male fish is widely used as an indicator of endocrine disruption. We studied the induction of vitellogenin mRNA in liver from several species of fish, both maintained in fish tanks or captured in the wild. Our procedure requires minute amounts of liver samples (down to 50 mg), and can be applied to field samples if the appropriate RNA-stabilisation agent is used. We used reverse-transcriptase PCR and quantitative real-time PCR for detection and precise quantitation of vitellogenin mRNA levels. Male mummichog (Fundulus heteroclitus) exposed to 17 beta-estradiol contained levels of vitellogenin mRNA up to 30 times higher than in untreated females and treatment with nonylphenol resulted in a weak but consistent induction of this transcript. We also studied levels of vitellogenin mRNA in a population of common carp (Cyprinus carpio) from the Anoia river, a river known for its high levels of estrogenic alkylphenols. The results were consistent with recorded data for fish from this sampling site. Finally, we also detected vitellogenin mRNA in barbs (Barbus graellsi), a species for which no vitellogenin sequence was available. The use of mRNA quantitation techniques for analysis of feral and cultured fish of different species opens the possibility of more precise detection and further control of the noxious effects of contaminants on the local fauna exposed to them.

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Section: Discussionsupporting

confidence: 84%

Section: Introductionmentioning

confidence: 99%

Use of vitellogenin mRNA as a biomarker for endocrine disruption in feral and cultured fish

García-Reyero¹,

Raldúa

Quirós

et al. 2004

Analytical and Bioanalytical Chemistry

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“…The presence of VTG in crustacean males may be extremely useful for future studies on vitellogenesis regulation, which has yet to be resolved for crustacean species. VTG in male crustaceans may also serve as a biomarker for xenobiotic compounds in the environments, as is the case in some fish species [73,74].…”

Section: Discussionmentioning

confidence: 98%

Relationship Between Vitellogenin and Vitellin in a Marine Shrimp (Penaeus semisulcatus) and Molecular Characterization of Vitellogenin Complementary DNAs1

Avarre

Michelis

Tietz

et al. 2003

Biology of Reproduction

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The source of yolk proteins in crustacean ovaries has been the subject of controversy for several decades, and both extraovarian and intraovarian synthesized proteins have been implicated. To offer a new insight into the relationship of vitellogenin (VTG) and vitellin (VT), a comparison of extraovarian VTG and ovarian VT of the marine shrimp Penaeus semisulcatus was performed at the protein and cDNA levels. Two cDNAs (7920 and 2068 nucleotides [nt]) were sequenced for VTG from the ovary and one cDNA (7920 nt) was sequenced from the hepatopancreas. VTG cDNA from hepatopancreas was similar to VTG cDNA from ovary. Although a VTG gene was also found in the males, approximately 7.8-kilobase transcripts were only detected in the ovary and hepatopancreas of females. The mRNA expression pattern was related to the stage of ovarian development and to the molt cycle, as determined by real-time polymerase chain reaction assay. VTG and VT apoproteins were composed of two and three major subunits, respectively, as shown by SDS-PAGE. N-terminal sequences of these subunits revealed the presence of a cleavage site at a consensus motif for a subtilisin-like endoprotease in VTG and VT and an additional cleavage site in VT revealed by an unidentified endoprotease. These results indicate that penaeid shrimps constitute a unique model for vitellogenesis, showing intraovarian gene expression and synthesis of yolk protein.

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“…After electrophoresis, the gels were stained for 30 min in 1 L of 1ϫ TAE (40 mM Tris acetate, 2 mM Na 2 -ethylenediaminetetraacetic acid [pH 7.4]) containing SYBR Green I nucleic acid gel stain (Molecular Probes, Eugene, OR, USA) per manufacturer's recommendation with no destaining step. Gels were scanned digitally on a FluorImager 595 (Molecular Dynamics, Sunnyvale, CA, USA) and resultant images were analyzed with ImageQuant (Molecular Dynamics) software as described in a previous investigation [33].…”

Section: Pcr and Gel Assay Protocolmentioning

confidence: 99%

17α-ethynylestradiol-induced vitellogenin gene transcription quantified in livers of adult males, larvae, and gills of fathead minnows (Pimephales promelas)

Lattier

Reddy

Gordon

et al. 2002

Environmental Toxicology and Chemistry

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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 17alpha-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 Competimers 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.

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Vitellogenin gene transcription: A relative quantitative exposure indicator of environmental estrogens

Cited by 39 publications

References 45 publications

Use of vitellogenin mRNA as a biomarker for endocrine disruption in feral and cultured fish

Use of vitellogenin mRNA as a biomarker for endocrine disruption in feral and cultured fish

Relationship Between Vitellogenin and Vitellin in a Marine Shrimp (Penaeus semisulcatus) and Molecular Characterization of Vitellogenin Complementary DNAs1

17α-ethynylestradiol-induced vitellogenin gene transcription quantified in livers of adult males, larvae, and gills of fathead minnows (Pimephales promelas)

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