Biology Project Abstract
CHANGES IN GLUTAMATE TRANSPORTER MRNA EXPRESSION AND PROTEIN LEVELS FOLLOWING LIGHT DAMAGE TO MOUSE RETINA (2003)
Presenter:
Neelima Vidula, Illinois Mathematics and Science Academy, 1500 West Sullivan Road, Aurora, IL, 60506; vneelima@imsa.edu
Mentors:
Mr. V.Joseph Dudley, Northwestern University Medical School, Department of Ophthalmology, 300 East Superior Avenue, Chicago, IL, 60611; 312-503-7503; vjdudley@northwestern.edu
Dr. Vijay Sarthy, Northwestern University Medical School, Department of Ophthalmology, 300 East Superior St., Chicago, IL, 60611; 312-503-7503; vjsarthy@northwestern.edu
Abstract:
Glutamate, a neurotransmitter, plays a role in numerous diseases of the central nervous system such as epilepsy, stroke, and Huntington disease. High concentrations of glutamate can damage the mammalian retina, leading to eye disorders such as glaucoma. GLAST and EAAT5, glutamate transporters, may prevent detrimental glutamate levels by controlling the uptake of excess glutamate. In order to elucidate the relationship between glutamate levels and retinal damage induced by long-term light exposure, BALB/c albino mice were exposed to constant light from 0 to 8 weeks. Every two weeks, the retinas of these mice were isolated, and the total RNA and protein were extracted from the retinal tissue. PCR experiments were then performed, using primers against GLAST and EAAT5. Retinal sections were cut on a cryostat to visualize the light damage. Immunocytochemistry experiments that detect the GFAP protein, which is increased in response to injury, were performed to confirm damage in the retina. Western Blotting experiments were conducted to detect EAAT5 in the retina. Data from PCR experiments against GLAST are being used to verify previous findings that GLAST mRNA levels decrease in the retina following long-term light exposure. Moreover, preliminary PCR results indicate that EAAT5 can be detected in the mouse retina.