Approximately ten million people across the globe have Parkinson's Disease (PD), a dopaminergic (DA) degenerative disease in the brain's substantia nigra. The cause and cure of PD remain a mystery, however heavy metals have been indicated as one cause. In PD symptom management, increases in dosage leads to side effects re quiring further medication to treat the side effects. As diet/supplemental treatment gains attention in the medical world, it may also be applied to PD patients to help reduce dosages to mitigate the detrimental side effects, like dementia. Caffeine's studied potential to increase lifespan, in journal article “Lifespan Extension Induced by Caf feine in Caenorhabditis elegans is Partially Dependent on Adenosine Signaling“, may apply to PD by protecting the DA neurons to help reduce the heavy reliance on medication.

Using the reliable neuroscience model organism Caenorhabditis elegans (C.elegans), I specifically tested caffeine's ability to protect the DA neurons from the heavy metal Cadmium (Cd). By utilizing the egls, 1 BZ555 C'elegans transgenic strain, the health/presence of DA could be measured through fluorescent imaging due to the Green Fluorescent Protein (GFP) spliced into the mutant nematode DA structures. Based on the previously refer enced article, the concentration of 5mM was used for caffeine as that dosage increased lifespan without significant effects on metabolic processes. I hypothesized with 5mM of caffeine, C.elegans DA structures will be protect ed against exposure to Cadmium.

Trials were conducted for four groups: Unexposed, 5mM Caffeine, 35uM Cd, 5mM + 35uM Cd. The con trolled environment was maintained with NGM agar and a dead bacteria lawn of K12 E.coli. Cadmium groups re ceived exposure through adding 0.5mL of a 35uM Cd solution from serial water dilutions; to control for the effect of moisture, 0.5mL of distilled water was added to the non Cd groups. Caffeine groups received caffeine through plating a 5mM concentration in bacteria broth; all group bacteria lawns were heat killed to prevent significant bac terial metabolization of caffeine. All nematodes were age synchronized through "Timed Egg Laying' or dissolving adult worms in a 20% alkaline hypochlorite solution and centrifuging the eggs out to plate. All neurons were im aged after a span of 8 days.

To collect data, worms were placed on microscope slides with 10.8mM sodium azide in M9 Buffer for an esthesia. Using a blue filter with the fluorescent light, the GFP in the DA structures actively fluorescence green and were imaged using Microscope 2.0 on a greyscale. Image) was then used to analyze images for a neuron value using the CTAF (Corrected Total Area Fluorescence = Intensity Density - (Area of Neuron)(Mean Back ground Fluorescence)] formula representing the amount of neuron present. Higher values reflected healthier neurons and the means of the groups were graphed for statistical comparison.

Using ANOVA, the means of the unexposed, 5mM Caffeine, and 35uM Cd were all significantly different from one another with a p value <<0.01. The 5mM Caffeine + 35uM Cd was significantly higher than the 35uM Cd with a p value << 0.5. The significance of the 5mM Caffeine + 35uM Cd being greater than the 35uM Cd concludes caffeine protects the DA structures from the degenerative Cadmium.

Future studies may further test exact concentration variations between caffeine and Cd as well as combina tions with current PD medications before being introduced in patient supplementation. With more precise technol ogy, such as CRISPR-Cas9, better target specific drugs may be created to stop the DA damage in PD patients with a reduction in harmful side effects.