Optic fissure (OF) and dorsal retina (DR) tissue collection and principle component analysis (PCA). (A) Sagittal section through the zebrafish eye at the level of the lens at 32, 48 and 56 hpf, representative of before, during and after optic fissure fusion. White dotted lines indicate the tissue (inferior is optic fissure and superior is dorsal retina) harvested for RNA-seq transcriptome analysis. Scale bar 50 µm. (B) PCA of RNA-seq transcriptome data expression patterns using regularized-logarithm transformed count data in DESeq2. Time (hours post fertilization, hpf) is represented by colour and origin of tissue by shape.

Failure of optic fissure fusion in ntn1a morphant zebrafish embryos. Expression levels (TPM, transcripts per million) of ntn1a candidate DEG showing temporal expression at 32,48 and 56 hours post fertilization (hpf) (A). (B) Wholemount and ocular morphology of 56 hpf wildtype (wt) and ntn1a morphant embryos, showing loss of ntn1a results in an ocular coloboma phenotype, scale bar 500 µm. (C) Histological analysis confirming loss of optic fissure fusion in ntn1a morphants. Scale bar 50 µm.