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| Status |
Public on Dec 01, 2022 |
| Title |
Symptomatic Domoic Acid exposure in zebrafish |
| Organism |
Danio rerio |
| Experiment type |
Expression profiling by array
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| Summary |
Domoic acid (DA) is a naturally produced neurotoxin synthesized by the marine diatom genus Pseudo-nitzschia. DA accumulates in filter-feeders such as shellfish, and can produce severe neurotoxicity when contaminated seafood is ingested. DA poisoning is a significant public health concern, and though seafood regulations have effectively minimized the human risk of severe acute poisoning, the effects of exposure at asymptomatic levels are poorly understood. The objective of this study was to determine the effects of exposure to symptomatic and asymptomatic doses of DA on gene expression patterns in the zebrafish brain. We exposed adult zebrafish to either a symptomatic (1.1 ± 0.2 μg /g) or an asymptomatic (0.31 ± 0.03 µg DA/g fish) dose of DA by intracelomic injection and sampled at 24, 48 and 168 h post-injection. Transcriptional profiling was done using Agilent and Affymetrix microarrays. Our analysis revealed distinct, non-overlapping changes in gene expression between the two doses. We found that the majority of transcriptional changes were observed at 24 hours post-injection with both doses. In response to symptomatic dose exposure, we observed 328, 5, and 1 genes were differentially expressed at 24, 48 and 168 hours post-injection, respectively. In contrast, 136, 12, and 28 genes were differentially expressed in the asymptomatic dose at the same time points. Pathway analysis revealed symptomatic DA exposure affected genes associated with pathways including cell adhesion, inflammation, and amyloid proteins. Among the pathways enriched among genes differentially expressed with asymptomatic exposure were circadian rhythms and endocrine signaling. Overall, these results suggest that transcriptional responses are specific to the DA dose and that asymptomatic exposure can cause long-term changes. Many of the differentially expressed genes are important players in neuronal, neuroimmune and neuroendocrine function. Further studies are needed to characterize the potential downstream neurobehavioral impacts of DA exposure.
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| Overall design |
Wild-type zebrafish (Danio rerio, AB strain), approximately five months of age, were obtained from Oregon State University (Sinnhuber Aquatic Research Laboratory Corvallis, OR). Fish were maintained at the Northwest Fisheries Science Center (NWFSC, Seattle, WA) in a ZebTec stand-alone recirculating and continuously-monitored zebrafish rack system with UV sterilizer (Techniplast, Exton, PA). Water temperature was maintained at 26°C, and fish were kept on a 12:12 hr light:dark cycle and fed daily with BioVita fish feed (Bio-Oregon, Longview, WA). Adult zebrafish were exposed to domoic acid (DA) or an identical volume of vehicle (PBS) via intracoelomic (IC) injection at one year of age. DA was purchased from Sigma Chemical Corp. (St. Louis, MO). The dose of DA used in this experiment was 1.1 ± 0.2 μg /g fish. Dose standard deviations represent slight differences in individual fish body weights. Injections were of 10μL volume delivered with 33-gauge needle in a custom-made auto-injector from Hamilton® (Reno, NV). After each injection, fish were observed for 30-45 minutes to note the instance of any neurobehavioral excitotoxic signs (e.g., circle- or spiral-swimming). This is considered to be a “symptomatic dose” as these fish showed behavioral symptoms soon after injection, which were ameliorated within five minutes. Each treatment consisted of 10-12 individual fish (biological replicates) per time period.
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| Contributor(s) |
Hidayat AS, Lefebvre K, MacDonald J, Bammler T, Aluru N |
| Citation(s) |
36198224 |
| Submission date |
Jun 19, 2020 |
| Last update date |
Mar 02, 2023 |
| Contact name |
James William MacDonald |
| E-mail(s) |
jmacdon@uw.edu
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| Organization name |
University of Washington
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| Department |
Environmental and Occupational Health Sciences
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| Street address |
4225 Roosevelt Way NE
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| City |
Seattle |
| State/province |
WA |
| ZIP/Postal code |
98105-6099 |
| Country |
USA |
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| Platforms (1) |
| GPL1319 |
[Zebrafish] Affymetrix Zebrafish Genome Array |
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| Samples (17)
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| Relations |
| BioProject |
PRJNA640534 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE152814_RAW.tar |
38.2 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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