Marcus Yaldo
Pronouns:
Research Mentor(s): Patrice Fort
Research Mentor School/College/Department: Ophthalmology / Medicine
Program: CG
Session: Session 1 (9:00am – 9:50am)
Authors: Marcus Yaldo, Zachary Sluzala, Patrice Fort
Abstract
Diabetic Retinopathy (DR) is a disease characterized by blurred, dark, or empty spots of vision caused by a combination of chronic retinal inflammation along with vascular and neuronal dysfunction. Currently, all treatment options target the late stages of DR which focus on inhibiting vascular endothelial growth, but do not cure the disease, only slow it down. There is still a dire need for early-stage treatments that focus on retinal neurodegeneration. aA-Crystallin are neuroprotective chaperone proteins expressed in the lens and retina, which have been shown to be upregulated in human donors and animal models with neurodegenerative diseases. T148 phosphorylation on aA-crystallin has been shown to be associated with their neuroprotective role. During DR the increase in aA-crystallin expression is accompanied by a concomitant decrease in T148 phosphorylation, which suggests that this phosphorylation is vital for the protein to carry out its protective role. To better understand the neuroprotective role of aA-crystallin, retinal neurons were grown and transfected to express either the Wild Type (WT) or the mutant forms which included T148A, the non-phosphorylatable, and T148D the phosphomimetic. The cells were then cultured in media with or without fetal bovine serum as a model of metabolic stress associated with cell death. Following culture, the cells were stained using Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) to allow for the visualization of cell death. The stained coverslips were then imaged by capturing microscope images of each part of the coverslip and stitching them together and then TUNEL-positive cells were counted. The hypothesis was that the T148A form and the Empty Vector (EV)(control) would show similarly high rates of cell death while the WT and T148D show relatively low cell death. Unfortunately, the stress condition tested showed very low induction of cell death preventing proper assessment of the neuroprotective role of aA-crystallin and the role of its phosphorylation. Further experiments will be aimed at testing various stress conditions (different duration or types of stress) to identify a stress condition that allows for testing this hypothesis.
