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Title: Development of quantitative real-time PCR assays for fathead minnow (Pimephales promelas) gonadotropin ß subunit mRNAs to support endocrine disruptor research.

Abstract

Fathead minnows (Pimephales promelas) are one of the most widely-used small fish models for regulatory ecotoxicology testing and research related to endocrine disrupting chemicals (EDCs). In this study, we isolated and sequenced cDNAs for fathead minnow follicle-stimulating hormone-like and luteinizing hormone-like β (FSHβ and LHβ) and glycoprotein α (GPα) subunits. Quantitative real-time PCR assays for measuring gonadotropin (GtH) β subunit transcripts were developed and used to examine “baseline” transcript levels over a range of age classes and reproductive states encompassed in EDC testing. In females, FSHβ and LHβ transcripts were greater in 4-5 month old than in younger fish and were significantly correlated with one another across all age classes examined. In males, FSHβ transcripts were greatest in 2-3 month old fish and were inversely correlated with various measures of testis development including, gonadal-somatic index (GSI), and histological stage. Overall, the pattern of GtHβ expression over age classes associated with gonad development was similar to that reported for other asynchronous-spawning fish. Despite significant changes in female GSI, gonad stage, and plasma vitellogenin within 24 h of spawning, GtHβ transcript levels in fish that had spawned within the preceding 24 h were not significantly different from those in fish that weremore » 2-3 days post-spawn and expected to spawn within the next 24 h based on spawning history. Results of this study provide insights related to the role of GtHs in fathead minnow reproductive development and function. Additionally they provide useful “baseline” data needed to design and interpret effective experiments for studying direct and indirect effects of EDCs on GtH subunit mRNA expression, which will facilitate a greater understanding of integrated system-wide responses of the fathead minnow brain-pituitary-gonadal axis to stressors including EDCs.« less

Authors:
; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
985075
Report Number(s):
PNNL-SA-47802
Journal ID: ISSN 1532-0456; TRN: US201016%%1760
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Resource Relation:
Journal Name: Comparative Biochemistry and Physiology Part C: Toxicology and Pharmacology, 145(2):171-183; Journal Volume: 145; Journal Issue: 2
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; DESIGN; FATHEAD MINNOW; FEMALES; GLYCOPROTEINS; GONADOTROPINS; GONADS; MALES; PLASMA; TESTING; fathead minnow; endocrine disruptors; PCR

Citation Formats

Villeneuve, Daniel L., Miracle, Ann L., Jensen, Kathleen M., Degitz, Sigmund J., Kahl, Michael D., Korte, Joseph J., Greene, Katie J., Blake, Lindsey S., Linnum, Ann, and Ankley, Gerald T.. Development of quantitative real-time PCR assays for fathead minnow (Pimephales promelas) gonadotropin ß subunit mRNAs to support endocrine disruptor research.. United States: N. p., 2007. Web. doi:10.1016/j.cbpc.2006.11.003.
Villeneuve, Daniel L., Miracle, Ann L., Jensen, Kathleen M., Degitz, Sigmund J., Kahl, Michael D., Korte, Joseph J., Greene, Katie J., Blake, Lindsey S., Linnum, Ann, & Ankley, Gerald T.. Development of quantitative real-time PCR assays for fathead minnow (Pimephales promelas) gonadotropin ß subunit mRNAs to support endocrine disruptor research.. United States. doi:10.1016/j.cbpc.2006.11.003.
Villeneuve, Daniel L., Miracle, Ann L., Jensen, Kathleen M., Degitz, Sigmund J., Kahl, Michael D., Korte, Joseph J., Greene, Katie J., Blake, Lindsey S., Linnum, Ann, and Ankley, Gerald T.. Thu . "Development of quantitative real-time PCR assays for fathead minnow (Pimephales promelas) gonadotropin ß subunit mRNAs to support endocrine disruptor research.". United States. doi:10.1016/j.cbpc.2006.11.003.
@article{osti_985075,
title = {Development of quantitative real-time PCR assays for fathead minnow (Pimephales promelas) gonadotropin ß subunit mRNAs to support endocrine disruptor research.},
author = {Villeneuve, Daniel L. and Miracle, Ann L. and Jensen, Kathleen M. and Degitz, Sigmund J. and Kahl, Michael D. and Korte, Joseph J. and Greene, Katie J. and Blake, Lindsey S. and Linnum, Ann and Ankley, Gerald T.},
abstractNote = {Fathead minnows (Pimephales promelas) are one of the most widely-used small fish models for regulatory ecotoxicology testing and research related to endocrine disrupting chemicals (EDCs). In this study, we isolated and sequenced cDNAs for fathead minnow follicle-stimulating hormone-like and luteinizing hormone-like β (FSHβ and LHβ) and glycoprotein α (GPα) subunits. Quantitative real-time PCR assays for measuring gonadotropin (GtH) β subunit transcripts were developed and used to examine “baseline” transcript levels over a range of age classes and reproductive states encompassed in EDC testing. In females, FSHβ and LHβ transcripts were greater in 4-5 month old than in younger fish and were significantly correlated with one another across all age classes examined. In males, FSHβ transcripts were greatest in 2-3 month old fish and were inversely correlated with various measures of testis development including, gonadal-somatic index (GSI), and histological stage. Overall, the pattern of GtHβ expression over age classes associated with gonad development was similar to that reported for other asynchronous-spawning fish. Despite significant changes in female GSI, gonad stage, and plasma vitellogenin within 24 h of spawning, GtHβ transcript levels in fish that had spawned within the preceding 24 h were not significantly different from those in fish that were 2-3 days post-spawn and expected to spawn within the next 24 h based on spawning history. Results of this study provide insights related to the role of GtHs in fathead minnow reproductive development and function. Additionally they provide useful “baseline” data needed to design and interpret effective experiments for studying direct and indirect effects of EDCs on GtH subunit mRNA expression, which will facilitate a greater understanding of integrated system-wide responses of the fathead minnow brain-pituitary-gonadal axis to stressors including EDCs.},
doi = {10.1016/j.cbpc.2006.11.003},
journal = {Comparative Biochemistry and Physiology Part C: Toxicology and Pharmacology, 145(2):171-183},
number = 2,
volume = 145,
place = {United States},
year = {Thu Mar 01 00:00:00 EST 2007},
month = {Thu Mar 01 00:00:00 EST 2007}
}