Abstract

Parkinson's disease (PD) is a well known disease estimated to affect 2% of people above age 60. It is a progressively debilitating disease affecting movement and motor control resulting in a steadily decreasing quality of life. PD can progress for as long as 20 years and does not always result in death but has been shown to significantly shorten life expectancy as well as increase the likelihood of death caused by choking, falls, and pneumonia. The most widely accepted cause is based on a lack of dopamine (DA) in the motor control centers of the brain. The current treatments focus on replacing the naturally occurring DA as well as extending the life of endogenous DA. A new approach focused on and preventative treatment is being developed. This new approach involves antagonism of a receptor in DA-producing neurons. Antagonism of this receptor results in higher levels of DA production and lessens the effects of PD. A number of antagonists have been identified and it has also been shown that the use of some common antagonists in food has been correlated with a reduced incidence of PD.

Down-Regulation of A2A Receptor as a Possible

Treatment for Parkinson’s DiseaseBen Demuth, Maleka Hashmi Ph.D, Department of Biology,

University of Wisconsin Stout, Menomonie WI 54751Methodology

Stiatal Neural Cell CultureStriatal cells were seeded and cultured in a medium prepared by Genlantis specifically for neural growth, from seven to thirty days in a 37 °C 1-5% CO2 incubator. The culture medium contained varying amounts of an A2A antagonist, caffeine. Cell confluency was monitored by phase-contrast microscopy; the cells were characterized using cell surface or intracellular markers and checked for contamination.

Liquid Chromatography/ Mass Spec In future experiments, two sets of cells will be grown; one will act as the control with just growth medium, while the other set will be grown with caffeine in its growth medium. Samples of both growth media will be periodically extracted and analyzed for DA levels.   ImmunohistochemistryIn future experiments, the viability of the striatal cells will be determined by IHC. This will entail fixation of the cells, application of primary antibody for typical neuronal markers, application of secondary antibody and visualization of protein of interest after color development with 0.03% diaminobenzidine.

References

Ibbotson, T, & Goa, K L (2002). Management of Parkinson's Disease The Role of Entacapone. Disease Management Health Outcomes, 10, 644-654.Brudin P, Li J & Holton JL.(2008). Research in motion: the enigma of Parkinson's disease pathology spread. Nature Reviews Neuroscience, 9, 741-754.Cilia R, et al. (2006). "Long-term Efficacy of Entacapone in Patients with Parkinson's Disease and Motor Fluctuations - A Six-Year Clinical Follow-Up StudyHack SP, Christie MJ (2003). "Adaptations in adenosine signaling in drug dependence: therapeutic implications". Critical Reviews in Neurobiology 15 (3-4): 235–74.

Results/ Discussion

Preliminary data suggest high susceptibility of culture to stress, contamination, and supplementation. Commercially coated culture flasks provide inadequate cell adhesion.

Proliferation possible using Fibroblast Growth Factor and Epidermal Growth Factor. Differentiation possible using Fetal Bovine Serum.

Rat E18 Striatal Differentiation FBS Response. p2, 13 days

Rat E18 Striatal Differentiation FBS Response p2, 20 days

Future Goals

Continue Culture with Caffeine supplementation.

Quantify A2a protein, DA

Animal Models