Excess glucose or fat differentially affects gene expression and appetite regulation during zebrafish embryogenesis

Zebrafish embryos were exposed to 3% glucose (24-72 hpf) and injected 2 nL of BODIPY-FL C12:0 dissolved in canola oil directly into the yolk sac at 24 hpf. At 72 hpf embryos were collected for analysis. Quantitative PCR analysis of glucose exposed embryos demonstrated that glucose exposure activates the insulin pathway while FFA/TAG injection activates the lipolysis and beta-oxidation pathways. At 72 hpf embryos were collected and bulk RNAseq was performed on control, glucose exposed and FFA/TAGs injected embryos. Embryos were treated with 3% glucose or injected into yolk sac with BODIPY-labeled fluorescent lipid dissolve in canola oil solution (1 mg/ml) at 24 hpf. At 72 hpf, samples were flash frozen. 10 embryos were used per condition with a total of six replicates (60 embryos total) for our experimental groups: glucose exposed or FFA/TAG injected embryos. A total of five replicates were used for the control group. Zebrafish embryos were sent to the UMMC Molecular and Genomics Core Facility (www.umc.edu/genomicscore) for RNA isolation, QC, and bulk RNAseq.