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The electroretinogram (ERG) is a powerful approach for investigating visual function in zebrafish ocular disease models. However, complexity, cost, and a literature gap present as significant barriers for the introduction of this technology to new zebrafish laboratories. Here, we introduce a simplified and effective method to obtain zebrafish ERGs. In-house assembled recording electrodes and a custom 3D-printed platform were used to gather high-quality and consistent ERG data from zebrafish at 3 developmental timepoints—larval, juvenile, and adult. Fish were tested under both scotopic (dark-adapted) and photopic (light-adapted) conditions to differentiate between the rod and cone systems, respectively. Robust ERG waveforms across all developmental timepoints were obtained using the methodology presented here. We observed an overall increase in signal amplitude as development progressed, reflecting maturation of the zebrafish retina. Oscillatory potentials could also be isolated from the generated waveforms. This simplified approach to the zebrafish ERG can generate waveforms comparable to the existing approaches and helps reduce barriers for zebrafish laboratories studying ocular development and disease.

The electroretinogram (ERG) is a powerful approach for investigating visual function in zebrafish ocular disease models. However, complexity, cost, and a literature gap present as significant barriers for the introduction of this technology to new zebrafish laboratories. Here, we introduce a simplified and effective method to obtain zebrafish ERGs.In-house assembled recording electrodes and a custom 3D-printed platform were used to gather high-quality and consistent ERG data from zebrafish at 3 developmental timepoints-larval, juvenile, and adult. Fish were tested under both scotopic (dark-adapted) and photopic (light-adapted) conditions to differentiate between the rod and cone systems, respectively.Robust ERG waveforms across all developmental timepoints were obtained using the methodology presented here. We observed an overall increase in signal amplitude as development progressed, reflecting maturation of the zebrafish retina. Oscillatory potentials could also be isolated from the generated waveforms.This simplified approach to the zebrafish ERG can generate waveforms comparable to the existing approaches and helps reduce barriers for zebrafish laboratories studying ocular development and disease.

Purpose Analysis of photoreceptor morphology and gene expression in mispatterned eyes of zebrafish growth differentiation factor 6a (gdf6a) mutants. Methods Rod and cone photoreceptors were compared between gdf6a mutant and control zebrafish from larval to late adult stages using transgenic labels, immunofluorescence, and confocal microscopy, as well as by transmission electron microscopy. To compare transcriptomes between larval gdf6a mutant and control zebrafish, RNA-Seq was performed on isolated eyes. Results Although rod and cone photoreceptors differentiate in gdf6a mutant zebrafish, the cells display aberrant growth and morphology. The cone outer segments, the light-detecting sensory endings, are reduced in size in the mutant larvae and fail to recover to control size at subsequent stages. In contrast, rods form temporarily expanded outer segments. The inner segments, which generate the required energy and proteins for the outer segments, are shortened in both rods and cones at all stages. RNA-Seq analysis provides a set of misregulated genes associated with the observed abnormal photoreceptor morphogenesis. Conclusions GDF6 mutations were previously identified in patients with Leber congenital amaurosis. Here, we reveal a unique photoreceptor phenotype in the gdf6a mutant zebrafish whereby rods and cones undergo abnormal maturation distinct for each cell type. Further, subsequent development shows partial recovery of cell morphology and maintenance of the photoreceptor layer. By conducting a transcriptomic analysis of the gdf6a larval eyes, we identified a collection of genes that are candidate regulators of photoreceptor size and morphology.