Dynamic changes occurring in living cells can be accessed in vitro by measuring the concentrations of cell-specific metabolites released into media bathing the cells. Dopamine-producing neuronal cell cultures release both dopamine and a metabolite, homovanillic acid, into the culture media. Fluctuations in the levels of DA and of HVA can be correlated to survival or death of these cells. The purpose of this study was to determine DA-HVA turnover rate using High Performance Liquid Chromatography (HPLC) in a neuroblastoma cell line (SH-SH5Y). The concentration of HVA ranges from 0.1 to 1 µg/mL in the feeding media of these cells, while dopamine concentrations ranged between 1 – 5 µg/mL. Previous studies have shown 100 to 1000 fold higher concentrations of both of these substances in similarly treated dopamine-producing cell lines. In conclusion, either the equipment currently available at Cornell is not sensitive enough to adequately detect the levels of HVA and DA in the media bathing cell cultures or the SH-SY5Y cell line does not release these substances and is therefore not an appropriate model of dopaminergic neurons.
Jessica Allison, ’04 Springfield, IL
Majors: Biochemistry & Molecular Biology and Chemistry
Sponsor: Barbara Christie-Pope