222— Identifying Regeneration Negative Zebrafish

Zebrafish are a useful model organism in studying developmental biology. One topic of interest is their ability to regenerate tissues, including retinal cells. Uncovering genetic pathways for this disease could be applicable to humans, as humans and zebrafish have numerous analogous genes. Scientists use mutants with non-functioning genes to elucidate where genes lie in a pathway. We studied mutants, which are suspected to lack the regenerative abilities of wild type zebrafish; the specific gene is unknown. Our goal was to confirm our family of fish as regenerative mutants. First, they were dark adapted and then placed in intense light for three days. We examined the retinas for regeneration under a fluorescence microscope. If they are regenerating they should be brightly expressing GFP (protein that glows under fluorescent light), but fail to express bright GFP if they are not. The eyes were surgically removed and cryosectioned. From here, the slides will be further examinedunder a confocal microscope in order to see if the antibodies bind to lower numbers of retinal cells in our mutant line compared with normal fish. We then plan on repeating this experiment for the other zebrafish in the same line as these parent zebrafish.