1) The document discusses how chronic alcohol intake may interact with genetic risk factors for Parkinson's disease (PD) and affect brain function. 2) It explores how heterozygous mutants of the DJ-1 gene, which is involved in oxidative stress response and implicated in familial PD, may influence sensitivity and tolerance to ethanol in Drosophila melanogaster. 3) The results suggest that different DJ-1 mutants may play roles in initial sensitivity, tolerance development, and behavioral sensitization to ethanol exposure as well as synaptic plasticity.

1. 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 genetic factors in Parkinson’s disease
Emily Park1, Jessica Burciaga, Erick Saldes, and Kyung-An Han
1Neuroscience 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 Dj-1α/+ Dj-1β/+ Dj-1α/+;
Dj-1β/+
Fluorescence(au)
Male: Basal Fluorescence
Combined Gamma Lobe
0
1
2
3
4
5
Canton-S Dj-1α/+ Dj-1β/+ Dj-1α/+;
Dj-1β/+
Fluorescence(au)
Female: Basal Fluorescence
Combined Gamma Lobe
0
0.5
1
1.5
2
2.5
Canton-S Dj-1α/+ Dj-1β/+ Dj-1α/+; Dj-1β/+
Fluorescence(au)
Male: Lateral Gamma Lobe
0X 6X
*
DJ-1α/+ DJ-1β/+ DJ-1α/+; DJ-1β/+
0
0.5
1
1.5
Canton-S Dj-1α/+ Dj-1β/+ Dj-1α/+; Dj-1β/+
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 worldwide1.
• Symptoms: resting tremors, bradykinesia, and postural instability1.
• Only 10% of cases are familial, while 90% are idiopathic1.
• Oxidative stress is a key pathogenesis process2.
• Cellular exposure to volatile reactive oxygen species (ROS)2.
• DJ-1 is a recessive PD gene involved in oxidative stress response3.
Dopamine (DA)
• PD involves a loss of dopaminergic neurons in the substantia nigra1.
• Neurotransmitter involved in addiction, movement, reward, learning and
memory.
• Dopamine precursors L-DOPA is a hallmark medication for PD2.
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 activity4.
• Causes oxidative stress5.
• Chronic alcohol induces adaptive
changes in brain activity that involve
dopamine:
• Tolerance
• Behavioral sensitization
• Dependence4.
• Flies have most of the PD genes, including DJ-1α and DJ-1β6.
• 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’-lobeα-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 Dj-1α/+ Dj-1β/+ Dj-1α/+; Dj-1β/+
Female Sedation Profile
DJ-1α/+ DJ-1β/+DJ-1β/+ DJ-1α/+; DJ-1β/+
MST, mean sedation time
Tolerance
0
10
20
30
40
50
60
70
Canton-S Dj-1α/+ Dj-1β/+ Dj-1α/+; Dj-1β/+
ToleranceIndex(%)
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)
1ST anti-DLG antibody made in a mouse
2ND 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
DJ-1α/+ DJ-1β/+
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/+ Dj-1β/+ Dj-1α/+; Dj-1β/+
Male Sedation Profile
DJ-1α/+ DJ-1β/+ DJ-1α/+; DJ-1β/+
0
5
10
15
20
25
CS Dj-1α/+ Dj-1β/+ Dj-1α/+; Dj-1β/+
MST(min)
*
*
Female Initial Sensitivity
DJ-1α/+; DJ-1β/+DJ-1α/+ DJ-1β/+
0
5
10
15
20
25
CS Dj-1α/+ Dj-1β/+ Dj-1α/+; Dj-1β/+
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
Malesengagedincourtship(%)
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
CSFemaleCSMale
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/+