Investigation of endoplasmic reticulum stress markers in A kainic acid formed excitotoxicity model

SARI A. Aydın Adnan Menderes University Institute of Health Sciences Medicine Faculty Clinical Biology Program Master Thesis, Aydın, 2020 Endoplasmic Reticulum (ER) stress occurs as a result of the accumulation of misfolded or unfolded proteins. ER stress is an adaptive stress response observed in almost all brain diseases, cancer and metabolic diseases. Excitotoxicity occurs when excess glutamate accumulates in synapses. Kainic acid is a toxin that leads to excitotoxicity in cellular and animal models. Kainic acid realated excitotoxicity causes stress in the cell. Therefore, our aim in this study is to investigate the relationship between ER stres and excitotoxicity by investigating two major ER stress markers ATF5 and phosphoryl eIF2. MTT Assay Test was used to determine the cell viability with the administration of 50, 200, 1000 µM doses of kainic acid 24 hours. After 24 hour incubation, protein was extracted both from control and kainic acid treated cells. The protein concenration was determined by Bradford Assay and the expression of ATF5 and phosphorylated eIF2was determined by western blot method. It was shown for the first time that ATF5 and phosphorylated eIF2 levels in neublastoma cells did not change after kainic acid treatment. As a result of our study, we thought that 50, 200, 1000 µM kainic acid treatment for 24 hours is not enough to induce Endoplasmic Reticulum stress measurable by ATF5 and phosphorylated eIF2. Increased concentration or duration of kanic acid treatment or the use of different markers could be tried in further experiments to induce ER stress. Key words: ATF5, Phosphoryl eIF2, Excitotoxicity, Endoplasmic Reticulum Stress, Kainic Acid