Effect of EDTA on CalModulin 1 (CAM1) Gene Expression and Cell Viability in Tetrahymena thermophila
The use of metal ion chelators, such as EDTA, is ever present in many of our daily products including shampoo, body soaps, and conditioners. For this project, we wanted to explore how the presence of EDTA influences the viability of Tetrahymena thermophila and expression of the Calmodulin 1 (CAM1) gene. We hypothesized that if EDTA is present in the media, CAM1 expression would increase but Tetrahymena viability would decrease. The CAM1 gene codes for the CalModulin 1 protein which requires calcium for activation and is involved in many cellular processes such as the Phospholipase C signaling pathway and intracellular movement. To test our hypothesis, Tetrahymena cells were cultured either in media containing no EDTA or in a media with 5mM EDTA for 24 hours. During the 24-hour treatment period, Tetrahymena cells from both groups were monitored. The cell viability was measured using a hemocytometer four different times throughout the 24-hour growth period. Immediately following the 24 hours, RNA extraction and Reverse Transcription quantitative-Polymerase Chain Reaction (RT-qPCR) were performed to measure CAM1 gene expression. The presence of EDTA was predicted to increase CAM1 gene expression and decrease cell viability due to the calcium sequestering properties of EDTA, which would decrease the amount of calcium available for CalModulin 1 activation.