1. Stress as a Non-Genetic Cause
for Bipolar Disorder
Presenters:
Anant Naik, Department of Biomedical Engineering
Bhavani Murakonda, Department of Biomedical Engineering
Advised by:
Dr. Atsushi Asakura, Department of Neurology
University of Minnesota, Twin Cities

2. The Fundamentals
of Bipolar Disorder
Grande et al., 2015, The Lancet
Life Progression of Bipolar Disorder
Mania
Hypomania
Euthymia
Subthreshold Depression
Major Depression
Mixed State
Severityof
Mania
Severityof
Depression Macro-level Background

3. The Fundamentals
of Bipolar Disorder
Micro-level Background
Cell Density Deficits in the
Prefrontal Cortex
Sagittal View Coronal View
Ongur et al., 1998, PNAS

4. The Fundamentals
of Bipolar Disorder
Cellular-level Background
Cell Density Deficits in the Prefrontal
Cortex
Ongur et al., 1998, PNAS
Glial Number in PFC for Mood Disorders
15
10
5
x106
GlialNumber
fBD - Familial Bipolar Disorder
oBD - Other Bipolar Disorder

5. The Relation Between
Stress and Anxiety
Is Bipolar Disorder abnormally prevalent in patients exposed to
situations with chronic stress?Q.
Posing the Question
Iraq and Afghanistan war
veterans are more likely to have
Anxiety and Mood disorders,
particularly Bipolar Disorder.
Bagalman, 2013, Congressional
Research Service

6. The Relation Between
Stress and Anxiety
Is Bipolar Disorder abnormally prevalent in patients exposed to
situations with chronic stress?Q.
Childhood
Trauma
Sexual &
Physical
Abuse
Combat
Veterans
YES

7. The Relation Between
Stress and Anxiety
Is Bipolar Disorder abnormally prevalent in patients exposed to
situations with chronic stress?Q.
Sample Characteristics Sample Size Rate of PTSD%
Bipolar patients admitted for mania or mixed 71 17
National general population survey: respondents with bipolar I
characterized by euphoria, grandiosity, and excessive energy 29 39
Inpatient and outpatient bipolar patients 50 40
Bipolar patients, manic or mixed: first admission for psychosis 77 21
Bipolar I and II outpatients recruited from community 288 7
Bipolar patients: first admission for psychosis 102 11
Bipolar I and II, treatment-seeking outpatients in the Systematic Treatment
Enhancement Program for bipolar disorder 475 17
Bipolar I and II, treatment-seeking outpatients 122 19
Otto et al., 2004, Bipolar Disorder

8. Our Hypothesis
Macro-level Hypothesis
Endocrine GC
Feedback to
stress
Endocrine
response to
stress
GC release due
to stress
PFC
Stress
Hypothalamus
Adrenal Gland
Cerebrum and Cerebellum
1. Chronic Stress induces an
upregulation in the
endocrine response
2. High concentrations of
glucocorticoids (GC) are
released
3. Cells in the Prefrontal
Cortex are degenerated in
the long run

9. Prefrontal Cortex
Our Hypothesis
Cellular-level Hypothesis
Neuron
Astrocyte
Blood vessel
GC
Neuron
Blood vessel
GC
Degeneration
of Astrocyte
In Prefrontal Cortex
Blood Vessel
Neuron
Astrocyte
GC
Astrocytes in the Prefrontal Cortex absorb glucocorticoids (GC),
resulting in their degeneration, and thus Bipolar Disorder.
Time
Neuron
Astrocyte
Blood vessel
GC
Neuron
Blood vessel
GC
Degeneration
of Astrocyte
In Prefrontal Cortex
Blood Vessel
Neuron
Degeneration of
Astrocyte
GC
Blood ves
Neuron
Blood ves
GC
Degeneration
of Astrocyte
Bipolar Disorder
Bipolar Disorder

10. Astrocytes Degenerate
with GC exposure in vitro34 K Unemura et al
results suggest that glucocorticoids inhibited astrocyte GR knockdown led
Fig. 2. Corticosterone (CRT) and dexamethasone (DEX) reduced the number of astrocytes
not induce astrocytic damage. A, B: The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazol
formed 72 h after treatment with CRT (0.01 – 1 μM) and DEX (0.01 – 1 μM). C: The lacta
performed 72 h after treatment with CRT (1 μM) and DEX (1 μM) or 24 h after treatment w
assay was performed 72 h after treatment with CRT (1 μM); DEX (1 μM); RU486 (0.3 – 3 μ
(10 μM), a mineralcorticoid receptor antagonist. n = 4. ***P < 0.001 vs. control, ###
P < 0.0
significant.
These results suggest that secretion of high concentra-
tions of glucocorticoids induced by repeated ACTH ad-
tion (Figs. 2 and 3). T
those of a previous rep
F
k
t
e
s
t
C
a
p
B
t
0
Cortisone Impact on
Astrocyte Number
Cortisone Impact on
GR RNA Prevalence
Cortisone Impact on
Astrocyte Functionality
MTTreductionofactivity
%ofcontrol
Control 0.01 0.1 1
Corticosterone (µM)
%ofBrdU+/GFAP+cells
%ofBrdU+/GFAP+cells
Unemura et al., 2012, J Phar Sci.

11. Astrocytes Degenerate
with GC exposure in vivo
Unemura et al., 2012, J Phar Sci.
corticotropic hormone (ACTH) administration decreased glucocorticoid receptor (GR) expression and
in vivo. A, B: GR expression in the frontal cortex (A) and hippocampus (B) after 14 days of saline or
e bottom graphs show the quantified data of the GR/GAPDH ratio. n = 3. C: GFAP and GAPDH were
ng in the frontal cortex after 14 days of repeated ACTH administration. D: Neuronal nuclei (NeuN)- or
stained by immunohistochemistry and their numbers calculated from 30 slices. E: Representative pic-
Saline ACTH
GFAP
P
NumberofGFAP+cells
P < 0.05
Saline ACTH
ACTH – Adrenocorticotropic Hormone
ACTH vs. Saline Injections in Mice
Prefrontal Cortex
Aggregate Astrocytes for
mPFC and Hippocampus

12. Astrocytes Degenerate
with GC exposure in vivo
Cerqueira et al., 2007, J Neurosci
Volume of Neurons in mPFC: Pre/post-stress
* (P < 0.05), ** (P < 0.01)

13. Significance
Current Inclusion/Exclusion Criteria
 Familial History
 Behavioral Analysis
Appropriate Diagnosis
of Bipolar Disorder
x Familial History
 Behavioral Analysis
Misdiagnosis
Total Patients with
Bipolar Disorder

14. Significance
Impacting Inclusion/Exclusion Criteria
 Familial History
 Behavioral Analysis
Appropriate Diagnosis
of Bipolar Disorder
 Behavioral Analysis
x Familial History
Misdiagnosis
 Chronic Stress History
 Behavioral Analysis
Total Patients with
Bipolar Disorder

15. Proposed Experiments
Behavioral
Tests
Immuno-
staining
Stem Cell
and Gene
Therapy
Measure decline in
glial densities by
exposure to chronic
stress in naive mice
Correlate Decline in
glial density to
behavioral
progression in mice
Promote astrocyte
growth in PFC using
stem cell therapy in
naïve and Bipolar
mouse models

16. Thank You
Questions? Concerns? Comments?
Acknowledgements:
For their guidance for the project, we would like to thank
Dr. Atsushi Asakura, Ph.D., Department of Neurology
Dr. Russell Carter, Ph.D., Department of Neuroscience