Role of Stiffness Markers in Astrocytes in Glaucoma Pathology

Glaucoma is a disease that increases pressure within the eye which applies stress to the optic nerve, leading to blindness. Most research has focused on different cells in the optic nerve, but this research focuses on a glial cell called astrocytes, which stabilizes cell connection, maintains the eye’s immune status, and maintains nutrient levels. Previous research has shown that stiffness around the optic nerve can be indicated by proteins which get upregulated. This research focuses on these proteins (Fibronectin, Collagen IV, ɑ-Smooth Muscle Actin) in an astrocyte, therefore, this project focuses on trying to understand the role of these stiffness markers. It was hypothesized that the levels of these proteins will change in response to glaucoma injury. This was tested by culturing astrocytes from a human cadaver’s optic nerve for 3-5 days in Astrocyte Growth Media. Once the cells reached 90% confluency, they were placed into separate chamber slides, then starved with serum-less DMEM. Half of the slides were placed into hypoxic conditions while the rest underwent normal conditions. These cells were then observed under a microscope at 4h, 16h, and 24h. Immunocytochemistry was then done on these cells for the three markers. The preliminary results indicate that healthy astrocytes do express all three markers, indicating potential for glaucomatous studies. Unfortunately, astrocytes could not survive hypoxic conditions as they died even in 4 hours. These preliminary studies enhance our understanding of stiffness markers that may play a role in glaucoma as compared to healthy astrocytes.