254 -- Fluorescent In-Situ Hybridization in Zebrafish with the Neurod4 Gene

Zebrafish are used as a model organism within experimental retinal research due to the developmental process of their eyes being similar to that of humans. This developmental process consists of apoptosis triggering the muller glial cells within the retina causing proliferation of progenitor cells. Similarities between the retinal development of zebrafish and that of humans, has led to the further exploration of the transcription factor neurod4 which may be of importance throughout the process of neuronal development. Not much is known about neurod4 but it is hypothesized to have a correspondingly important impact on human neuronal development. Due to a lack of research around neurod4, this study looks to analyze how the expression of the neurod4 transgene zebrafish compares to the expression of the endogenous gene. Endogenous gene expression can be visualized through the use of green fluorescent protein (GFP) as its absorption of blue light results in emittance of GFP that allows for visualization. It is hypothesized that the transgene will share the same activity as the promoter found on neurod4, showing that the transgene is able to exhibit the same expression as the endogenous gene. Additionally, if the endogenous gene’s function is knocked down, the cell should not be able to stimulate new growth as it is deleterious to the cell. Labeling of the neurod4 gene can be observed through Fluorescence in situ hybridization (FISH) which will be implemented in this study. FISH will allow detection of mRNA presence and its expression.