Evaluating Alzheimer's Disease Therapeutics on mitoNEET Expression

Alzheimer's disease (AD) is a neurodegenerative disease that affects over 5 million Americans. The disease is characterized by the formation of senile plaques of the amyloid beta and neurofibrillary tangles within the brain that can impair the patient's memory and behavior. These symptoms of AD develop slowly and worsen over time. Currently there is no known cause or cure for AD, therefore treatment is restricted to alleviating symptoms. A new approach to AD focuses on mitochondrial dysfunction, which is when the mitochondria release reactive oxidative species that cause damage and changes to the expression of tissues, proteins, and genes. MitoNEET is a newly discovered mitochondrial protein that is thought to regulate bioenergetics in cells. The focus of our research is to help resolve the mechanism of AD by evaluating mitoNEET as a potential target for treatment. Fluorescence microscopy is used to evaluate changes in protein expression. This was used to assess changes in mitoNEET-GFP when exposed to current AD therapeutics. One treatment is isoproterenol, which is a bronchodilator that has been shown to upregulate mitoNEET. Our preliminary studies verify that isoproterenol upregulates the expression of mitoNEET in N2a cells after a 24-hour exposure. The results show a two-fold increase in the relative integrated density when exposed to 1, 10, and 100 uM of isoproterenol. We did not detect a significant difference in relative integrated density of mitoNEET-GFP when exposed to donepezil hydrochloride (5, 50, or 500 μM), rivastigmine L-tartrate (0.5, 5, 50, 500 μM), or galantamine hydrobromide (0.1, 1, 10, 100 μM). Further studies will investigate mitoNEET regulation in response to oxidative stress.