Pilot testing and optimization of a larval fathead minnow high throughput transcriptomics assay

RNA-seq expression assays were done in larval Pimephales promelas (Fathead minnow, FHM) exposed to 10 chemicals (3 metals, 4 Neonicotinoids, and 3 pharmaceuticals) over a range of concentrations to derive relatively short term transcriptomics based points of departure (tPODs). tPODs were compared to apical points of departures (aPODs) to investigate the potential use of the FHM tPODS as part of a high throughput approach that might be used in a regulatory context. Comparative gene expression profiling analysis of RNA-seq data from exposed larval FHM; untreated controls vs. half-log dilution series of treatment concentrations (n=11) to estimate tPODs for ten chemicals (CuSO4, NiSO4, ZnSO4, Clothianidin, Flupyradifurone, Imidacloprid, Thiacloprid, Fluoxetine, Paroxetine and Sertraline). For each treatment chemical, 96 well plate with individual 24 hour old FHM larva in each deep (1 mL) well. 8 replicates at each concentration, 24 hour static exposure, followed by phenotypic (survival, behavior) observation and subsequent homogenization and RNA sequencing of fish in doses that did not experience excessive mortality.