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| Status |
Public on Jul 30, 2010 |
| Title |
Investigations of gene expression in fathead minnow (Pimephales promelas) brain tissue reveal toxicological impacts of RDX exposure |
| Organism |
Pimephales promelas |
| Experiment type |
Expression profiling by array
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| Summary |
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.
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| Overall design |
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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| Contributor(s) |
Gust K, Wilbanks MS, Guan X, Pirooznia M, Habib T, Yoo L, Wintz H, Perkins E |
| Citation missing |
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| Submission date |
Mar 31, 2010 |
| Last update date |
Mar 22, 2012 |
| Contact name |
Kurt A Gust |
| E-mail(s) |
kurt.a.gust@usace.army.mil
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| Phone |
601-634-3593
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| Organization name |
US Army ERDC
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| Department |
Environmental Laboratory
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| Lab |
Environmental Genomics and Systems Biology Team
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| Street address |
3909 Halls Ferry Rd.
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| City |
Vicksburg |
| State/province |
MS |
| ZIP/Postal code |
39180 |
| Country |
USA |
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| Platforms (1) |
| GPL10277 |
US Army ERDC Fathead minnow brain tissue cDNA 4K Microarray |
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| Samples (30)
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| Relations |
| BioProject |
PRJNA126597 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE21153.pdf.gz |
49.8 Kb |
(ftp)(http) |
PDF |
| GSE21153_RAW.tar |
14.3 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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