Naumchuk Y. Biotechnological basis of a lactate dehydrogenase system for testing glioprotective compounds in optic nerve head astrocytes

Optic nerve head astrocytes (ONHAs) are the major glia cell type in the non-myelinated optic nerve head where they contribute to extracellular matrix synthesis. Pathological changes in glaucoma include reactive astrocytosis, a process characterized by altered astrocyte gene and protein expression and extracellular matrix remodeling. ONHAs are highly sensitive to mechanical and oxidative stresses and crucial for the maintenance of retinal ganglion cell physiology and function. Therefore, glioprotective strategies with the goal to preserve and/or restore the structural and functional viability of ONHAs to slow progression of glaucoma are of high clinical relevance. Available methods for the evaluation of drug candidates with glioprotective potential in ONHAs are poorly developed, laborious and costly, thus causing a challenge for the development of the novel glioprotective strategies. Therefore, the objective of this work was to establish the scientific premise for glioprotection in primary ONHA culture and to develop a biotechnological method for the screening of glioprotective compounds based on their efficacy in reducing or preventing the deleterious effects of oxidative stress in ONHAs. We formulated the following five specific aims to reach our objective: 1) optimization of the isolation and culturing techniques for primary rat ONHAs; 2) development and validation of a lactate dehydrogenase (LDH) release assay for ONHAs; 3) optimization of chemically-induced oxidative stress to use as an insult in primary rat ONHAs; 4) evaluation of the effects of oxidative stress and hyperbaric pressure on ONHAs; 5) validation of the glioprotective screening platform using antioxidants in primary rat ONHA culture.