Visual impairments are widespread in the population due to common retinal ailments such as macular degeneration, diabetic retinopathy, and visual decline with age. Unfortunately, humans lack the ability to regenerate damaged retina tissue and therefore, research exploring gene regulation in regenerative animals can provide important insight to combat retinal diseases. At day four of embryonic development, the chicken is able to regenerate the neural retina through reprogramming of the retinal pigment epithelium (RPE) by responding to the signaling protein FGF2. However, by day five this regeneration ability is lost. The molecular mechanisms underlying the decline in regenerative capacity are not yet understood. In order to understand the processes associated with RPE injury and regeneration, we performed RNA-sequencing on chicken RPE following injury and FGF2 treatment at days 4 and 5 of development. This method allows for the quantification of total gene expression; resultant data was processed through the use of RStudio and the DESeq2 package. Using this method, we identified a novel signaling pathway, HGF-MET, as being upregulated during retina regeneration in the presence of FGF2. To elucidate the role of HGF-MET signaling, we inserted a bead containing HGF into the chick eye following retina removal to see if HGF is able to induce retina regeneration in the absence of FGF2. Our findings showed that HGF on its own is insufficient to induce retina regeneration. Future studies will focus on describing the interaction between FGF2 and HGF signaling to determine their respective roles in regeneration, as well as contrasting the injury responses observed in E4 and E5 RPE. Specifically, this research has allowed me to refine laboratory techniques and understand how tissue regeneration is tightly linked with gene regulation, which is integral to my pursuit of a dual graduate / medical degree in tissue engineering.
Author: Alexandra Danciutiu
Faculty Advisor: Katia Del Rio-Tsonis, Biology
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