Gene expression profiling of zebrafish embryos at 1 hour post injection of PAMPs

We use the zebrafish embryo model to study the transcriptome responses to flagellin and Pam3CSK4. Therefore, we injected these PAMPs into the caudal vein at the 27 hours post fertilization and took samples at 1 hour post injection. Overall design: This deep sequence study was designed to determine the gene expression profile by Pam3CSK4 and flagellin injection. RNA was isolated from embryos at 1 hour post injection. Wildtypes and tlr2- and tlr5a- morphants zebrafish embryos were micro-injected into the caudal vein with 1ng of Pam3CSK4, 0,1 ng flagellin , or water as a control at 27 hours post fertilization. After injections embryos were transferred into fresh egg water and incubated at 28°C. At 1 hour post injection triplicates of 10 to 15 embryos per condition were snap-frozen in liquid nitrogen, and total RNA was isolated using TRIZOL reagent.

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Tudorache C, Slabbekoorn H, Robbers Y, Hin E, Meijer JH, Spaink HP, Schaaf MJM