Methamphetamine (METH) is a highly addictive psychostimulant that has been demonstrated to produce toxicity in brain tissue with the loss of some neurons. Therefore, METH can be used as a pharmacological tool to induce neurodegeneration in the brain. Our laboratory has been investigating the mechanism whereby METH induces neural damage in the striatum. Recently, it has been demonstrated that striatal neuropeptides mediate this METH induced damage. Some neuropeptides are pro and others are anti-damage. This project focuses on a natural product, green tea, which has been shown by our laboratory to abrogate METH-induced neural damage.

1. Effects of Epigallocatechin Gallate
on Astrocyte Activation In METH-
induced Toxicity
Columbia University
2015 SPUR Fellow at Hunter College
Christopher Lites

2. Epidemiology of Methamphetamine (METH)
● 15 - 16 million users worldwide (UNODC, 2007)
● Second most widely abused drug after
cannabis (UNODC, 2007)
● Major public health concern
● 5.8% of Americans (>12) have used at least
once (Substance Abuse and Mental Health Services
Administration, 2007)
(Substance Abuse and Mental Health Service
Administration, 2007)

3. Larger Implications of Examining METH toxicity
● Dangers of METH include symptoms of certain neurodegenerative diseases
● Long-lasting damage to dopaminergic neurons mimics process of
Parkinson’s Disease
● METH is an ideal toxic candidate to produce an animal model for
Parkinson’s disease
(McCann et. al, 1998)

4. What does METH do to the brain?
METH
● Causes accumulation of Dopamine,
Serotonin and Norepinephrine in their
respective synaptic clefts
● Damages Mitochondria and proteins
● Produces reactive oxygen species (ROS)
● Damages synaptic function
(Yamamoto, 2005)

5. METH toxicity and Astrocyte Activation
● Astrocytes are glial cells that
respond to injury in the brain
● METH toxicity increases
astrocyte activation
● Astrocytes have inflammatory
properties that act as a
detriment normal brain function
METHNo METH
(Wilhelmsson, vol. 103 no. 46)

6. METH-Induced Astrocyte Activation in the Striatum
● Selective vulnerability in neuronal subpopulations for the striatum of
METH-injected mice
● Astrocytes are activated in METH injected mice
● Astrocytes produce neuroinflammation which exacerbates oxidative
damage

7. Epigallocatechin gallate (EGCG)
● Most abundant polyphenol (antioxidant) found in green tea
● Vast epidemiology data indicate a negative correlation between
occurrence of neurodegenerative disorders and green tea consumption
● Known to protects against METH-induced striatal apoptosis

8. Hypothesis
● We hypothesized that EGCG would protect from METH-
induced toxicity by attenuating activation of
astrocytes
o GFAP, a protein expressed by activated astrocytes,
will exhibit lower levels in mice that have been
injected with EGCG in addition to the METH

9. Establishing a Mouse Model of METH toxicity
● CD1 male mice
● 9 mice were assigned each of the four groups: saline (control), M30, M40,
and M50
o i.e M30 = 30mg of METH per Kg (body weight) CD1 Male Mice
● Mice were sacrificed 3 days post injection
● TUNNEL assay
● Confocal microscopy

10. TUNEL Assay of Cell Damage Resulting From Various METH Concentrations
Saline
(control) M30 M40 M50

11. M30 Dosage Exhibits Optimal Temperature Spike

12. Experimental Procedure
● Four groups were prepared
a. CD1 male mice injected with EGCG and M30
b. CD1 male mice injected with EGCG
c. CD1 male mice injected with M30
d. CD1 male mice injected with saline
● Mice were sacrificed 3 days after injections and their striata were
dissected
● Protein quantification
● Western Blot to identify GFAP levels in the varying groups

13. Expected Results
● GFAP levels
> >CD1 male mice
injected with
METH (M30)
CD1 male mice
injected with
METH (M30) and
EGCG
CD1 male mice
injected with
saline
CD1 male mice
injected with
EGCG
Greatest Least

14. Future Studies
● How does EGCG affect astrocytes? Does it have an indirect or direct
effect?
● Does EGCG block or ameliorate other mechanisms involved in METH
toxicity?
o Free radical production
o Lipid Peroxidation
o Activated microglia
o Activated pro-/anti- apoptotic proteins
● Does EGCG have an effect on enzymes that break down superoxide
dismutase (a radical scavenging enzyme)?

15. Acknowledgements
● Christine Gonzalez - Program Supervisor
● Allen Pan - Grad Student
● Dr. Jesus Angulo - Lab Supervisor and Mentor
○ M.S., Ph.D