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Transcriptomic points of departure calculated from rainbow trout gill, liver, and gut cell lines exposed to methylmercury and fluoxetine

ABSTRACT: The primary objective of this study was to determine if quantitative tPOD values could be derived from chemical exposure studies that followed OECD Test No. 249 (rainbow trout gill cell line) and expanding the assay to rainbow trout liver and intestinal epithelial cell lines. The secondary objective was to determine if tPOD values could be derived from similar studies on rainbow trout liver and gut cell lines. Methylmercury and fluoxetine were the test chemicals. From the resulting data, we compared tPOD data with literature-derived apical BMDs, characterized mechanisms of action through analysis of the transcriptomic data, and compared responses (cytotoxicity and transcriptomic) across the three cell types. By bringing together transcriptomics measurements and tPOD calculations with the OECD Test, and expanding the number of cell lines, we believe that this work can help establish a cost effective in vitro test method that can yield quantitative tPOD values that are protective of in vivo concentrations associated with adverse outcomes while also advancing knowledge of a test chemical’s mechanism of action.

ORGANISM(S): Oncorhynchus mykiss

PROVIDER: GSE205562 | GEO | 2022/06/13

REPOSITORIES: GEO

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Project description:Whirling disease, caused by the pathogen Myxobolus cerebralis, afflicts several salmonid species. Rainbow trout are particularly susceptible and suffer high mortality rates. The disease is persistent and spreading in hatcheries and natural waters of several countries, including the U.S., and the economic losses associated with whirling disease have been extremely large. In this study, gene expression profiling was conducted for resistant and susceptible rainbow trout strains following pathogen exposure with the primary objective of identifying specific genes associated with whirling disease resistance. Several genes were significantly up-regulated in skin following pathogen exposure for both the resistant Hofer and susceptible Trout Lodge rainbow trout strains. For both strains, response to infection appears to be associated with the interferon system. Metallothionein is differentially expressed between the resistant and susceptible strains and is a good candidate for future studies due to its diverse functional roles in inflammation and immune response. The present study has provided the first examination into the genetic basis underlying rainbow trout’s immune response and resistance to the whirling disease pathogen. The identified genes have allowed us to gain initial insight into the molecular mechanisms associated with a salmonid host’s immune response and resistance to whirling disease infection. Keywords: Disease state comparison. 1st comparison: exposed vs control. 2nd comparison: resistant vs susceptible strains Resistant (Hofer) and susceptible (Trout Lodge) rainbow trout strains were exposed to 2,000 triactinomyxons (whirling disease pathogen) per fish. Unexposed controls for each strain were treated identically to exposed other than their lack of pathogen exposure. After twenty-four hours, caudal fin tissue was collected for RNA extraction. Amplified RNA was competitively hybridized (exposed versus control) for each strain. Four biological replicates were used for each experimental condition and a balanced block design was incorporated.

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Transcriptomic points of departure calculated from rainbow trout gill, liver, and gut cell lines exposed to methylmercury and fluoxetine

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