Effects of agouti overexpression in the brain and skin transcriptome of zebrafish

Twenty transgenic fish overexpressing agouti-signaling protein (ASIP, Ceinos et al., Pigment Cell and Melanoma Research 28:196-209, 2015) and wild type (WT) zebrafish were placed in 6-liter tanks and acclimated during 30 days prior to sampling to minimize physiological differences. Animals were sacrificed and the brain+pituitary and the skin dissected under the stereo microscope, placed in trizol and stored at -80C until RNA extraction. Six biological replicates (3 females and 3 males) of each tissue (skin and brain) of each group (ASIP vs WT) were analyzed in a total of 6 slides containing 4 microarrays in the 4 × 44 k format. Global gene expression analyses were performed to search for both sex-dependent and transgene-dependent effects as well as the interaction between both factors (sex x phenotype) on the brain+pituitary and skin transcriptome. Wild type (Tu) or transgenic zebrafish overexpression agouti-signaling protein (ASIP) were reared in similar conditions. Brain+pituitary and skin were sampled from 6 ASIP (3 males + 3 females) and WT (3 males + 3 females) zebrafish. Samples were individually hybridized in a total of 6 slides containing 4 microarrays in the 4 × 44 k format. Microarrays included 60-mer probes to the unique transcripts from Ensembl and Unigene which were annotated by functional categories of GO and pathways of KEGG with bioinformatics package STARS