Project description:We have employed whole genome microarray expression profiling as a discovery platform to identify genes with differential expression in WT and hif1α mutants and consequent different response to normoxia or hypoxia conditions. Two different conditions were considered: 50 hpf mutants and WT in normoxia, 50 hpf mutants and WT after hypoxia chamber treatment (2 hours after 3% O2)
Project description:RNAseq was performed on hcar1-4+/+ and hcar1-4-/- zebrafish larvae with (2 h and 4 h) or without (0 h) Pseudomonas aeruginosa (PA) ear infection. The transcriptome profile generated here reveals PA14 infection-induced differential gene expression patterns between hcar1-4-/- and hcar1-4+/+ siblings.
Project description:After gastrulation, oviductal hypoxia maintains turtle embryos in an arrested state prior to oviposition. Subsequent exposure to atmospheric oxygen upon oviposition initiates recommencement of embryonic development. Arrest can be artificially extended for several days after oviposition by incubation of the egg under hypoxic conditions, with development recommencing in an apparently normal fashion after subsequent exposure to normoxia. To examine the transcriptomic events associated with embryonic arrestAfter gastrulation, oviductal hypoxia maintains turtle embryos in an arrested state prior to oviposition. Subsequent exposure to atmospheric oxygen upon oviposition initiates recommencement of embryonic development. Arrest can be artificially extended for several days after oviposition by incubation of the egg under hypoxic conditions, with development recommencing in an apparently normal fashion after subsequent exposure to normoxia. To examine the transcriptomic events associated with embryonic arrest, RNA-sequencing analysis was performed on embryos from freshly laid eggs and eggs incubated in either normoxia (oxygen tension ~159 mmHg) or hypoxia (<8 mmHg) for 36 hours (h) after oviposition (n = 5 per group). The patterns of gene expression differed markedly among the three experimental groups. Normal embryonic development in normoxia was associated with up-regulation of genes involved in DNA replication, the cell cycle, and mitosis, but these genes were commonly down-regulated after incubation in hypoxia. Many target genes of hypoxia inducible factors, including insulin-like growth factor binding protein 1, were down-regulated by normoxic incubation but upregulated by incubation in hypoxia. Notably, some of the transcriptomic effects of hypoxia in green turtle embryos resembled those reported by others to be associated with hypoxia-induced embryonic arrest in diverse taxa, including budding yeast (Saccharomyces cerevisiae), the annelid Caenorhabditis elegans, and zebrafish (Danio rerio). Thus, while among oviparous species, hypoxia-induced pre-ovipositional embryonic arrest appears to be a unique adaptation of turtles, mechanisms underlying hypoxia-induced embryonic arrest per se may be highly conserved across the diverse taxa in which this mechanism operates. We report the transciptional changes in Chelonia mydas embryos incubated in either normoxia or hypoxia comapred to freshly laid eggs.
