Figure 1

Mutation in pde6a zebrafish causes a premature stop codon. (A) Structure of zebrafish Pde6a protein with specific GAF (cGMP-specific phosphodiesterases, adenylyl cyclases and FhlA) and HDc [histidine (H) and/or aspartate (D) amino acid residues] domains. (B) Sanger sequencing identified the nucleotide mutation in exon 1 of pde6a sequence. Asterisk (*) denotes the substituted nucleotide in the knock-out pde6a sequencing trace. (C) A targeted fragment was amplified by PCR from the genomic DNA of adult zebrafish tail followed by an enzymatic digestion with Mwo1. The three rows present digested products for pde6a^Q70X, pde6a^WT, and pde6a^WT/Q70X fish. Mwo1 restriction site is shown on the side of the agarose gel image. The mutation is highlighted in bold. (D) Expression level of pde6a and pde6b mRNA revealed by qPCR in pde6a^Q70X and pde6a^WT zebrafish larvae. Expression of pde6a and pde6b were normalized using the zef1α reference gene. Error bars represent SD calculated from three replicas. ^****p < 0.0001.