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

Journal Article · Thu Mar 01 04:00:00 EST 2007 · Comparative Biochemistry and Physiology Part C: Toxicology and Pharmacology, 145(2):171-183

DOI:https://doi.org/10.1016/j.cbpc.2006.11.003· OSTI ID:985075

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

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.

• Research Organization: Pacific Northwest National Laboratory (PNNL), Richland, WA (US)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 985075
• Report Number(s): PNNL-SA-47802
• Journal Information: Comparative Biochemistry and Physiology Part C: Toxicology and Pharmacology, 145(2):171-183, Journal Name: Comparative Biochemistry and Physiology Part C: Toxicology and Pharmacology, 145(2):171-183 Journal Issue: 2 Vol. 145; ISSN 1532-0456
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
99 GENERAL AND MISCELLANEOUS
DESIGN
FATHEAD MINNOW
FEMALES
GLYCOPROTEINS
GONADOTROPINS
GONADS
MALES
PCR
PLASMA
TESTING
endocrine disruptors
fathead minnow