Gene expression in homozygous mbl/axin1 zebrafish

Homozygous masterblind (mbl-/-) zebrafish exhibit reduced or absent eyes and telecephalon, and the expansion of the diencephalic fates to the front of the brain. A missense mutation in the GSK3-binding domain of zebrafish axin1, a scaffolding protein in the Wnt signaling pathway, results in the mbl phenotype. In an effort to identify and study the genes affected by Wnt signaling, we used a 14,000-oligonucleotide-gene microarray to determine differentially expressed genes in mbl/axin1 (-/-) and wild type control zebrafish embryos and larvae. Keywords: zebrafish, Danio rerio, wild-type mbl, axin1, development RNA was isolated from pools of approximately 50 wild-type and from approximately 50 homozygous mbl/axin1 mutant zebrafish at 24, 48, and 72 hours post-fertilization (hpf). Two separate sets of embryos or 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) to identify genes affected in development in organisms with this mutation.