Project description:This study was performed to validate the newly developed CGP Atlantic cod 20K oligonucleotide microarray. Atlantic cod (Gadus morhua) received an intraperitoneal injection of either formalin-killed, atypical Aeromonas salmonicida (Asal) or PBS and transcriptional profiles of spleen tissues from Asal-injected fish were compared to those from pre-injection control fish and PBS-injected control fish. Gene expression profiles resulting from this study were compared to those from suppression subtractive hybridization library studies, that were previously performed on the same samples, and to literature. Gene expression patterns of single genes were confirmed by QPCR analysis. This study has shown that the newly developed CGP Atlantic cod 20K oligo microarray platform is a valuable tool for cod genomic research.
Project description:Lipid metabolism is essential in maintaining energy homeostasis in multicellular organisms. In vertebrates, the peroxisome proliferator-activated receptors (PPARs, NR1C) regulate the expression of many genes involved in these processes. Four Ppar subtypes from Atlantic cod (Gadus morhua) were recently cloned and characterized. However, the downstream regulatory role of Ppars in cod lipid metabolism is presently not well understood or described. Here we study the involvement of Atlantic cod Ppar subtypes in systemic regulation of lipid metabolism using the model agonists WY14,643, GW501516, and tetradecylthioacetic acid, employing a multiple omics approach after an in vivo exposure situation.
Project description:The aim of the exposure was to study the effects of activation of peroxisome proliferator-activated receptors (PPARs) in Atlantic cod (Gadus morhua), by injecting the fish with the compounds WY-14,643 and GW501516. Using luciferase reporter assay in vitro, we have shown that WY-14,643 activate Atlantic cod Ppara1 and Ppara2, while GW501516 activate Ppara1, Ppara2, and Pparb. The experimental set-up was as follows: Immature cod were injected at day 0 and day 4 with either high dose (40 mg/kg WY-14,643 and 4.0 mg/kg GW501516), low dose (4.0 mg/kg WY-14,643 and 0.4 mg/kg GW501516), or solvent control (10 % DMSO, 90 % teleost saline (2.41 mM KCl, 133.5 mM NaCl, 1.5 mM CaCl2, 0.79 mM MgSO4, 1 mM NaHCO3, 0.5 mM Na2HPO4)). At day 11, liver samples were collected from 9-12 male fish from each group (total of 50 samples). Total RNA was isolated from 50 mg of each sample using TRI reagent (Sigma), and 0.4 μg RNA were sequenced at the Genomics Core Facility at the University of Bergen on Illumina HiSeq 4000 (Illumina, Inc., San Diego, CA, USA).
