The effects of progesterone on the fathead minnow testis
Ontology highlight
ABSTRACT:
ORGANISM(S): Pimephales promelas
PROVIDER: GSE38288 | GEO | 2012/07/02
SECONDARY ACCESSION(S): PRJNA167622
REPOSITORIES: GEO
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Project description:The objectives of the study were to determine the effects of progesterone in the male testis. Reproductive fathead minnows were used in the study. Testis at stage 2 and 3 were dissected and testis explants were treated progesterone or no progesterone (10^-6M and 10^-8M) for 12 hours. Androgens were measured and there was a significant increase in testosterone, but not 11-tetotetosterone. Gene expression analysis was performed in the testis which genes were differentially regulated by progesterone. testis explants were treated with 10^-6M progesterone for 12 hours; 6 control and 4 treatment samples
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Project description:Metformin, along with its biotransformation product guanylurea, are commonly observed in municipal wastewaters and subsequent surface waters. Previous studies in fish have identified metformin as a potential endocrine active compound but there are inconsistencies in the literature with regard to effects. To further investigate the potential reproductive toxicity of metformin and guanylurea to fish, a series of experiments were performed with reproductively mature fathead minnows (Pimephales promelas). First, explants of mature fathead minnow ovary tissue were exposed to 0.001-100 µM metformin or guanylurea to investigate whether they can directly perturb steroidogenesis. Second, spawning pairs of fathead minnows were exposed to metformin (0.41, 4.1, 41 µg/L) or guanylurea (1.0, 10, 100 µg/L) for 23 d to assess impacts on reproduction. Lastly, male fathead minnows were exposed to 41 µg/L metformin, 100 µg/L guanylurea, or a mixture of both compounds, with samples collected over a 96 h time course to investigate potential impacts to the hepatic transcriptome or metabolome. Neither metformin or guanylurea effected estradiol or testosterone by ovary tissue exposed in vitro. In the 23 d exposure, neither compound significantly impacted transcription of endocrine-related genes in male liver or gonad, circulating steroid concentrations in male or female fish, or fecundity of spawning pairs. In the 96 h time course, 100 µg guanylurea/L elicited more differential gene expression than 41 µg metformin/L , and showed the greatest impacts after 96 h. A number of DEGs up-regulated after 24 h were subsequently down-regulated after 96 h, demonstrating time-dependent impacts of guanylurea on the liver. Overall, metformin and guanylurea did not elicit effects consistent with reproductive toxicity in adult fathead minnows at environmentally relevant concentrations. Where effects were identified using ‘omics approaches, guanylurea induced greater impacts than metformin.
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Project description:Fathead minnow ovary explants were incubated with DHT at 10^-6M for 6, 9, and 12 hours. Estrogen production was significantly increased in these ovaries at all three time points. control 6 hours (n=4), DHT 6 hours (n=5), control 9 hours (n=5), DHT 9 hours (n=5), control 12 hours (n=4), DHT 12 hours (n=4),
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Project description:Trenbolone and flutamide are prototypical model compounds for respectively androgen and anti-androgen modes of action. Trenbolone is an anabolic steroid used in cattle industry to increase weight gain and feed efficiency, and flutamide is a pharmaceutical used to treat prostate cancer. Androgens exert profound effects on the organization, development, and function of the male reproductive system through activation of androgen receptors (ARs). Flutamide, as an antiandrogen, was expected to temper the effects of trenbolone on androgen receptor (AR)-mediated gene expression. Female fathead minnows (Pimephales promelas) were exposed via the water to 0.05, 0.5 or 5 µg 17β-trenbolone/L; 50, 150 or 500 µg flutamide/L; or to a mixture of 0.5 μg/L trenbolone and 500 μg/L flutamide. After a 48 hr constant exposure gene expression in gonads was analyzed using a fathead minnow-specific 22,000-gene microarray. Trenbolone significantly changed the expression (P < 0.05) of roughly 750 different genes, while flutamide changed the expression of 1420 genes. Most of the genes that were regulated were distinct for the two chemicals. However, 70 genes were reciprocally regulated by the two treatments. Myelocytomatosis viral oncogene homolog (Myc), Yin Yang 1 (YY1) and interferon regulator factor 1 (IRF1) were among the important transcription factors reciprocally regulated by trenbolone and flutamide, suggesting they may be specifically regulated by the AR. These regulatory molecules, in conjunction with other reciprocally regulated transcription factors, may function as early molecular switches to control phenotypic changes in gonad tissue architecture and function. Fathead minnows exposed for 48h to 0.5 ug trenbolone, 500 ug flutamide, or a mixture of both.
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Project description:Production, usage and disposal of the munitions constituent (MC) cyclotrimethylenetrinitramine (RDX) has led to environmental releases on military facilities. The chemical attributes of RDX are conducive for leaching to surface water which may put aquatic organisms at risk of exposure. Because RDX has been observed to cause aberrant neuromuscular effects across a wide range of animal phyla, we assessed the effects of RDX on central nervous system (CNS) function in the representative aquatic ecotoxicological model species, fathead minnow (Pimephales promelas). A brain-tissue based cDNA library enriched for transcripts differentially expressed in response to RDX exposure was developed for fathead minnow and was transitioned to custom cDNA-based microarrays. All 4,128 cDNAs were sequenced, quality filtered and assembled yielding 3,018 unique sequences and 945 significant blastx matches (E ≤ 10-5). Bioassays were conducted exposing fathead minnows to RDX at 0.625, 1.25, 2.5, 5, 10 mg/L or an acetone-spike control for 10d. Overt toxicity of RDX in fathead minnow occurred only at the highest exposure concentration resulting in 50% mortality. Conversely, Bayesian analysis of microarray data indicated significant changes in transcript expression in fathead minnow brain tissue at RDX concentrations as low as 0.625 mg/L. In total, 154 microarray targets representing 44 unique transcript identities were differentially expressed in RDX exposures, the majority of which were validated by RT-qPCR. Investigation of molecular pathways, gene ontology and individual gene functions indicated that RDX exposures affected metabolic processes involved in: oxygen transport, neurological function, calcium binding / signaling, energy metabolism, cell cycle / cell proliferation, oxidative stress and ubiquitination. In total, our study indicated that RDX exposure affected molecular processes critical to CNS function in fathead minnow. 10 Day RDX Exposure, Brain Tissue Investigation: Sub-adult fathead minnows were exposed to RDX in a 10d dose-series experiment (0.625, 1.25, 2.5, 5.0, or 10 mg/L RDX) which included an acetone-spike control (1% acetone). Each experimental treatment included 8 replicate fish (48 total fish) and endpoints included mortality, total weight and neurotoxicogenomics. The 1.25mg/L dose was not included in the microarray experiment. Please see attached PDF file for detailed 'Balanced, Interwoven Loop Design'.
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