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Increased PD Risk PD Genetic Factor Chronic Ethanol Intake 1. Bonifati, V., Oostra, B. A., & Heutink, P. (2004). Linking DJ-1 to neurodegeneration offers novel insights for understanding the pathogenesis of Parkinson’s disease. Journal of molecular medicine, 82(3), 163-174. 2. Lev, N., Roncevich, D., Ickowicz, D., Melamed, E., & Offen, D. (2006). Role of DJ-1 in Parkinson's disease. Journal of molecular neuroscience, 29(3), 215-225. 3. Meulener, M., Whitworth, A. J., Armstrong-Gold, C. E., Rizzu, P., Heutink, P., Wes, P. D., ... & Bonini, N. M. (2005). Drosophila DJ-1 mutants are selectively sensitive to environmental toxins associated with Parkinson’s disease. Current Biology, 15(17), 1572-1577. 4. Bowirrat, A., & Oscar‐Berman, M. (2005). Relationship between dopaminergic neurotransmission, alcoholism, and reward deficiency syndrome. American Journal of Medical Genetics Part B: Neuropsychiatric Genetics, 132(1), 29-37. 5. Bondy, S. C. (1992). Ethanol toxicity and oxidative stress. Toxicology letters, 63(3), 231-241. 6. Attrill H, Falls K, Goodman JL, Millburn GH, Antonazzo G, Rey AJ, Marygold SJ; the FlyBase Consortium. (2016) FlyBase: establishing a Gene Group resource for Drosophila melanogaster. Nucleic Acids Res. 44(D1):D786-D792 7. Krashes, M. J., Keene, A. C., Leung, B., Armstrong, J. D., & Waddell, S. (2007). Sequential use of mushroom body neuron subsets during Drosophila odor memory processing. Neuron, 53(1), 103-115. Methods Flypub Assay Results Chronic alcohol intake and g enetic factors in Parkinson’s disease Emily Park 1 , Jessica Burciaga, Erick Saldes, and Kyung-An Han 1 Neuroscience Program, Wellesley College, Wellesley, MA Department of Biological Sciences, Neuroscience and Metabolic Disorders Project, University of Texas at El Paso, El Paso, TX Ethanol-induced Behaviors Immunohistochemistry Antennal Lobes Drosophila melanogaster Synaptic Molecule DLG Expression 0 1 2 3 4 5 Canton-S Fluorescence (au) Male: Basal Fluorescence Combined Gamma Lobe 0 1 2 3 4 5 Canton-S Fluorescence (au) Female: Basal Fluorescence Combined Gamma Lobe 0 0.5 1 1.5 2 2.5 Canton-S Fluorescence (au) Male: Lateral Gamma Lobe 0X 6X * DJ-1α/+ DJ-1β/+ DJ-1α/+; DJ-1β/+ 0 0.5 1 1.5 Canton-S Fluorescence (au) Female: Lateromedial Gamma Lobe 0X 6X * DJ-1α/+ DJ-1β/+ DJ-1α/+; DJ-1β/+ Ethanol (EtOH) Objective To investigate whether chronic alcohol intake interacts with genetic risk factors of Parkinson’s disease for brain functions. Parkinson’s Disease (PD) • Second most common progressive neurodegenerative disease worldwide 1 . • Symptoms: resting tremors, bradykinesia, and postural instability 1 . • Only 10% of cases are familial, while 90% are idiopathic 1 . • Oxidative stress is a key pathogenesis process 2 . • Cellular exposure to volatile reactive oxygen species (ROS) 2 . • DJ-1 is a recessive PD gene involved in oxidative stress response 3 . Dopamine (DA) • PD involves a loss of dopaminergic neurons in the substantia nigra 1 . • Neurotransmitter involved in addiction, movement, reward, learning and memory. • Dopamine precursors L-DOPA is a hallmark medication for PD 2 . This work is supported by: NIDA Award 4R25DA033613-05, NIH/NIAAA 1R15AA020996, NIH-NIMHD-RCMI Grant 2G12MD007592, and BBRC: Cytometry, Scanning and Imaging Core Facility. Mushroom body and fly pub schematics are courtesy of Ivan Mercado. IHC schematic courtesy of Paul Sabandal. In memory of Won Park, who taught me to ask questions incessantly and unapologetically. Acknowledgements • A legal drug, widely used drug. • Increases DA activity 4 . • Causes oxidative stress 5 . • Chronic alcohol induces adaptive changes in brain activity that involve dopamine: • Tolerance • Behavioral sensitization • Dependence 4 . • Flies have most of the PD genes, including DJ-1α and DJ-1β 6 . • In this study, we used heterozygous mutants DJ-1α/+ and DJ-1β/+ as well as the transheterozygous mutant DJ-1α/+;DJ-1β/+ to investigate their potential interactions with ethanol. -lobe α’-lobe α-lobe β-lobe β’-lobe The mushroom body of the fly brain has homology to the human hippocampus and is involved in behavioral plasticity. 7 L = Lateral gamma lobe, LM = Lateromedial, M = Medial, AIMPR = Anterior Inferior Medial Protocerebrum; Scale bar = 25μm 10 12 14 16 18 20 22 24 26 Exp 1 Exp 2 Exp 3 Exp 4 Exp 5 Exp 6 MST (min) CS Female Sedation Profile DJ-1α/+ DJ-1β/+ DJ-1α/+; DJ-1β/+ MST, mean sedation time Tolerance 0 10 20 30 40 50 60 70 Canton-S Tolerance Index (%) Male Female * * * DJ-1α/+ DJ-1α/+; DJ-1β/+ DJ-1β/+ p-value: * <0.05, ** <0.005, *** <0.0001 ns = not significant n = 6-12 To monitor synaptogenesis, we used immunostaining on the postsynaptic molecule DLG (PSD-95) 1 ST anti-DLG antibody made in a mouse 2 ND anti-mouse IgG conjugated with Alexa 488 DLG (PSD-95) Day of Adulthood 0 1 2 3 4 5 6 7 8 9 10 EtOH Exposure Day 1 2 3 4 5 6 MST Collect Rest Dissect & IH Disinhibition 10 12 14 16 18 20 22 24 26 Exp 1 Exp 2 Exp 3 Exp 4 Exp 5 Exp 6 MST (min) CS Dj-1α/+ Male Sedation Profile DJ-1α/+ DJ-1β/+ DJ-1α/+; DJ-1β/+ 0 5 10 15 20 25 CS MST (min) * * Female Initial Sensitivity DJ-1α/+; DJ-1β/+ DJ-1α/+ DJ-1β/+ 0 5 10 15 20 25 CS MST (min) DJ-1α/+ ns Male Initial Sensitivity DJ-1α/+; DJ-1β/+ DJ-1β/+ Behavioral Disinhibition and Sensitization 0 5 10 15 20 25 30 35 40 45 Exp 1 Exp 2 Exp 3 Exp 6 Males engaged in courtship (%) CS Dj-1α/+ Dj-1β/+ Dj-1α/+; Dj-1β/+ n = 6 DJ-1β/+ DJ-1α/+; DJ-1β/+ DJ-1α/+ Introduction Mean Sedation Time (MST) Disinhibition EtOH Record how long it takes for flies to sedate Score percentage of male flies that court other males Dissection, Immunohisto- chemical (IH) analysis CS Female CS Male No EtOH Exposure 6X EtOH Exposure NROI:AIMPR L LM M • Sedation profiles of Canton-S and all PD heterozygous mutants were comparable. • DJ-1β may be important for the initial sensitivity response to ethanol in females but not in males. • DJ-1α may be important for tolerance development to ethanol in females. • DJ heterozygous mutants tend to have reduced behavioral sensitization. • DJ heterozygous mutants may have reduced synaptic plasticity. Conclusions • Test additional ethanol exposures and concentrations to clarify the roles of DJ-1 in behavioral sensitization. • Examine additional synaptic molecules to uncover changes in synaptic plasticity. • Investigate an age factor in ethanol x gene interaction by testing aged flies. • Explore interaction of additional PD genes with ethanol. Future Direction References Oxidative Stress DJ-1α/+ DJ-1β/+ DJ-1α/+; DJ-1β/+ DJ-1α/+ DJ-1β/+ DJ-1α/+; DJ-1β/+
