1. Time-Dependent Activity
and Metabolic Markers
A Novel Characterization of
Ethanol Withdrawal
Anant Naik
IMPACT Program
Under the guidance of:
Dr. Doo-Sup Choi
Molecular Pharmacology and Experimental Therapeutics Track
Mayo Clinic, Rochester MN

2. Ethanol Withdrawal
Administration of
Alcohol Acute
Alcohol
Effects
Opposing
Neuroadaptation
Alcohol
tolerance
Removal of
Alcohol
Withdrawal
Syndrome
Recovery from
Neuroadaptation
CNS
Equilibrium
Alcohol-free
state
Littleton (1998) National Institute on Alcohol Abuse and Alcoholism
Himmelsbach Hypothesis

3. Administration of
Alcohol Acute
Alcohol
Effects
Opposing
Neuroadaptation
1
2
3
Various Models in Mice
- Vapor Chamber
- Oral Gavage
- Two Bottle Choice

4. Administration of
Alcohol Acute
Alcohol
Effects
Opposing
Neuroadaptation
1
2
3
Various Models in Mice
- Vapor Chamber
- Oral Gavage
- Two Bottle Choice
- Drowsiness
- Altered Mental Status

5. Administration of
Alcohol Acute
Alcohol
Effects
Opposing
Neuroadaptation
1
2
3
Various Models in Mice
- Vapor Chamber
- Oral Gavage
- Two Bottle Choice
- Drowsiness
- Altered Mental Status
Molecular Changes
GlutamateAdenosine
Mitochondrial Changes in
Astrocytes
- Migration
- Morphology
- Functionality

6. Alcohol
tolerance
Removal of
Alcohol
Withdrawal
Syndrome
4 New homeostasis
- Decreased effect of
EtOH
- Plasticity
56

7. Alcohol
tolerance
Removal of
Alcohol
Withdrawal
Syndrome
4 New homeostasis
- Decreased effect of
EtOH
- Plasticity
56
Time
BehavioralFrequency
Extinction
Phenomenon
Extinct Spontaneous
Relapse
Extinction
Start
Extinction
Burst

8. Alcohol
tolerance
Removal of
Alcohol
Withdrawal
Syndrome
4 New homeostasis
- Decreased effect of
EtOH
- Plasticity
56
Time
BehavioralFrequency
Extinction
Phenomenon
Extinct Spontaneous
Relapse
Extinction
Start
Extinction
Burst
Symptoms of Withdrawal
• Sleep difficulty
• Anxiety
• Loss of Appetite
• Nausea
• Rapid Heart Rate
• Lethargy
Medline, NIH

9. Can we identify objective time-dependent
markers for metabolic activity for ethanol
withdrawal?
Q
Lack of Metabolic
Considerations
Limited
Understanding of
Time-dependent
Changes
Insufficient
Biological Basis

10. Methods
Ethanol Dependence Schedule
Acute Injection
• Long Term Exposure
• BAC Measurement
Chronic
Exposure
Acute
Re-administration
6:00
18:00
Dark Cycle Light Cycle
10:00
Ethanol Exposure
No exposure
Outputs:
• Oximetry
• Activity
• Food IntakeVapor Chamber

11. Results
Blood Ethanol Concentration

12. Results
Long-term vs. Chronic Withdrawal
𝑅𝑒𝑠𝑝𝑖𝑟𝑎𝑡𝑜𝑟𝑦 𝑄𝑢𝑜𝑡𝑖𝑒𝑛𝑡 (𝑅𝑄) =
𝐶𝑂2 𝑒𝑙𝑖𝑚𝑖𝑛𝑎𝑡𝑒𝑑
𝑂2 𝑐𝑜𝑛𝑠𝑢𝑚𝑒𝑑A. B. Dark Cycle

13. Results
Long-term vs. Chronic Withdrawal
Dark Cycle
A. B.

14. Results
Re-administration of EtOH
A. B.

15. Results
Re-administration of EtOH
A. B. C.

16. Future Directions
• Pre-clinical comparisons
of genotype using
paradigm
• Comparison of RQ and
EE values with treatment
with Taurine or
Acamprosate during
alcohol withdrawal
Metabolite-based
Biomarkers of withdrawal,
observing glutamine,
adenosine, correlating
with metabolic behavior.
• Magnetic Resonance
Spectroscopy Imaging
• Micro-dialysis
Mitochondrial
Dysfunction and
regulation, size, density,
orientation, location in
neurons and astrocytes
Clinical monitoring of
Respiratory Quotient
and Energy expenditure
and correlates with
withdrawal of ethanol
1 2 3 4

17. Acknowledgements
Lab Members
Sierra Slade
Dr. David Hinton, Ph.D.
Phillip Starski
Wayne Olson
Dr. Ada Ho
Alfredo Oliveros
Katie Wininger
Sun Choi
Daniel Lindberg
Dr. Yun-Fang Jia
Dr. Paul Croarkin, D.O, M.S.
Dr. Doo-Sup Choi, Ph.D.
Summer Students
Betsey Daly
Joyce Yang
Erin Nosco
IMPACT Program
Dr. Katie Campbell
IMPACT Scholars
MPET
Dr. Mays
MPET Summer Students
“If I have seen further than others, it is by standing upon the
shoulders of giants.”
- Isaac Newton