1. Aubrey Hernandez Dr. Robert Haining Spring 2016
Competitive Binding of Parkinsonian-inducing Neurotoxins to Neuromelanin
Aubrey Hernandez
Dr. Robert Haining
Spring 2016

2. Parkinson’s Disease (PD)
7-10 million worldwide living with PD (Parkinson’s Disease Foundation, 2016)
1 million Americans (PD Foundation, 2016)
60,000 diagnosed each year (PD Foundation, 2016)
Age: 50+ (PD Foundation, 2016)
Symptoms: Tremors, bradykinesia, akinesia, cognition, emotions, postural instability (NIH, 2016)

3. Parkinson’s Disease (PD)
Death of dopaminergic neurons in substantia nigra
Dopamine can no longer reach the striatum
Causes problems with controlled movements
More specific on biology of PD
(By NIH)

4. Neuromelanin (NM) Dopaminergic neurons pigmented with neuromelanin
Hypothesized to be responsible for aiding in the controlled release of dopamine
How does NM tie in
(By BruceBlaus)

5. Research Questions Do neurotoxins interact directly with neuromelanin?
Can the interaction between neuromelanin and neurotoxins be disrupted by nicotine?
Is neuromelanin directly involved in the controlled release of dopamine?

6. Methodology NanoDrop 3300 (Thermo Scientific, 2013)
Measures relative fluorescence under either white, UV or blue light
RFU = Relative Fluorescence Unit
Source of Neuromelanin
Synthesized in the lab following protocol from Zecca’s Neuromelanin Interaction with Lipids and Peptides (Zecca, 2002)
Fe3+

8. 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)
Drug-induced Parkinson’s disease
Converted to MPP+
Dopaminergic uptake system/neuromelanin (D’Amato, 1986)
Nicotine
(By Harbin)

10. Effect of Altering [NM] on Interaction between MPTP and NM
Working on ways to quantitate it right now by the amount of iron it binds
Effect of Altering [NM] on Interaction between MPTP and NM

11. Nicotine Correlation between smokers and PD (Nefzger, 1968)
Neuroprotective effects (Singh, 2010)
(By Harbin)

15. Conclusion Presence of NM causes MPTP’s RFU to disappear
Indicates interaction
Presence of nicotine causes NM RFU to increase dramatically

16. Conclusion
Addition of nicotine to MPTP+NM solution causes a release of MPTP back into solution
Shows nicotine can disrupt the interaction between MPTP and NM
Addition of MPTP to nicotine+NM solution doesn’t cause a change in RFU
Shows MPTP cannot disrupt the interaction between nicotine and NM

17. Further Experimentation
MPP+ rather than MPTP
6-hydroxy dopamine (6-OHDA)
Chloroquine (D’Amato, 1987) and Caffeine (Singh, 2010)
Shown to have neuroprotective properties
Altering [Nicotine]
Further characterize interaction between dopamine and neuromelanin

18. References
Bush WD, G. J. (2006). “The surface oxidation potential of human neuromelanin reveals a spherical architecture with a pheomelanin core and a eumelanin surface”. PNAS,
By BruceBlaus. When using this image in external sources it can be cited as:Blausen.com staff. "Blausen gallery 2014". Wikiversity Journal of Medicine. DOI: /wjm/ ISSN Own work, CC BY 3.0,
By Harbin - Own work, Public Domain,
By Harbin - Own work, Public Domain,
By NIH - Public Domain,
D’Amato, Robert, Guillermo Alexander, Robert Schwartzman, et al. (1987). “Neuromelanin: A role in MPTP-induced neurotoxicity”. Life Sciences 40 (8):
D’Amato, ZP Lipman, and SH Snyder. (1986). “Selectivity of the parkinsonian neurotoxin MPTP: toxic metabolite MPP+ binds to neuromelanin”. Science 231(4741):
Di Monte, D.A and S.A Jewell. (2-14). “MPTP Neurotoxicity”. Encyclopedia of Neurological Sciences 2:
Dzierzega-Lecznar A, K. S. (2004). “GC/MS Analysis of Thermally Degraded Neuromelanin from the Human Substantia Nigra”. J Am Soc Mass Spectrom,
Liu Y, S. J. (2003). “Isolation and Biophysical Studies of Natural Eumelanins: Applications of Imaging Technologies and Ultrafast Spectroscopy”. Pigment Cell Res,
McGeer PL, McGeer EG. (2008). “Glial reactions in Parkinson’s disease”. Mov Disord 23,
Nefzger MD, Quadfasel FA, and Karl VC. (1968). “A retrospective study of smoking in Parkinson’s disease”. American Journal of Epidemiology 88:
Singh, Kavita, Seema Singh, Naveen Kumar Singhal, et al. (2010). “Nicotine and caffeine-mediated changes in gene expression patterns of MPTP-lesioned mouse striatum: Implications in neuroprotection mechanism”. Chemico-Biological Interactions 185 (2):
Thermo Scientific. (2013). “NanoDrop 3300 Fluorospectrometer”. Thermo Fisher Scientific Inc. Web.
W, C. (2004). “The Chemical Structure of Melanin”. Pigment Cell Res,
Wakamatsu K, F. K. (2003). “The structure of neuromelanin as studied by chemical degradative methods”. J Neurochem,
Zecca L, T. D. (2001). “Substantia nigra neuromelanin: structure, synthesis, and molecular behavior”. Mol Pathol,
Zecca, L. et al. (2002). “Interaction of Human Substantia Nigra Neuromelanin with Lipids and Peptides”. Journal of Neurochemistry 74 (4):
Zecca L, Z. F. (2006). “A proposed dual role of neuromelanin in the pathogenesis of Parkinson’s disease”. NEUROLOGY, S8-S11.