Transcriptome analysis of zebrafish yquem/urod mutant

Zebeafish yquem harbors a mutation in the gene encoding uroporphyrinogen decarboxylase (UROD), the fifth enzyme in heme biosynthesis, and was established as a vertebrate model for human hepatoerythropoietic porphyria (HEP). In an effort to investigate the unknown aspects of UROD deficiency pathogenesis, we used a 14,000-oligonucleotide-gene microarray to determine differentially expressed genes in yquem/urod (-/-) and wild type control zebrafish larvae. Keywords: zebrafish, Danio rerio, wild-type yquem, urod, porphyria RNA was isolated from pools of approximately 50 wild-type and from approximately 50 homozygous yquem/urod mutant zebrafish at 56, 72, 84 and 86 hours post-fertilization (hpf). One set of larvae were used for each time point. RNA from each replicate at each time point was separately reverse transcribed and labeled by direct incorporation with dUTP-Cy3 prior to hybridization to microarrays printed with zebrafish 50-mer oligonucleotide library (MWG/Ocimum) to identify genes affected in development in organisms with this mutation.