Development of Neuroprotection Assay and Analysis of Endogenous Production for Eicosanoyl-5-Hydroxytryptamide

Abstract

Parkinson’s Disease (PD) and Alzheimer’s Disease (AD) are neurodegenerative disorders that severely impact cognitive and motor function, eventually resulting in death. Both diseases are thought to be associated with the hyper-phosphorylation of the neuronal proteins α-synuclein and tau respectively. Epidemiological studies have noted a correlation between coffee consumption and reduced incidences of these diseases. This led to the identification of eicosanoyl-5-hydroxytryptamide (EHT) as a potential compound responsible for coffee’s neuroprotection as it was found to promote the methylation, and thus activity, of protein-phosphatase-2A (PP2A), which is thought to be responsible for the dephosphorylation of both tau and α-synuclein. Additionally, EHT has also been shown to be neuroprotective against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridin (MPTP), a neurotoxin that mimics PD. This work attempted to develop an assay for identifying endogenous EHT production as well as neuroprotection against glutamate excitotoxicity, as glutamate is thought to play a role in neurodegenerative disorders such as PD and AD. The development of a glutamate excitotoxicity assay with human SH-SY5Y neuroblastoma cells as well as PC12 rat adrenal medulla pheochromocytoma cells was unsuccessful due to the inability to achieve significant and cohesive cell death. However, the development of an assay for the identification of endogenous EHT production showed more promising results, as preliminary evidence for time and substrate dependence was found in addition to localizing activity to the membrane of the tissue extract. Further work is needed to confirm these results.