HARVARD BRAIN TISSUE RESOURCE CENTER
An NIH National Resource
A National Repository for the Acquisition and Distribution of High Quality
Postmortem Brain Tissue to the Neuroscience Community
Neuropathological Assessment
Microscopic Evaluation
Alzheimer’s Disease
Tangles
Distributive Diagnoses
Cases Collected 2002-March, 2007
Diagnosis
Intake
Distributive
Controls (Normal)
282
197
Alzheimer’s Disease
Alzheimer’s D., Lewy Body Variant
Diffuse Lewy Body Disease
Frontotemporal Dementia
411
356
21
38
30
Huntingtington’s Disease
Huntington’s Disease (Grades 3,4)
Huntington’s Disease (Grades 1,2
Huntington’s “at risk”
202
Parkinson’s Disease
Progressive Supranuclear Palsy
107
2
62
6
*Tourette’s Syndrome
11
11
Schizophrenia
Bipolar Disorder
Depression , Unipolar
*Autism
85
66
0
41
35**
34**
0
6
Total
114
59
2
1207
971
*Must get approval from appropriate tissue use committee. **Diagnosis pending on approximately 20
cases.
Standards of Practice
Brain banks must conform to the Uniform Anatomic Gift Act of 1987
Informed consent for brain donation is critical, particularly as
it relates to genomics research
Irreversible anonymization may eliminate ethical/legal issues
regarding genomics research.
Distribution of brain tissue is ethical when the tissue or its
bioderivatives are serving the common good
i.e. they help to identify the causes and new therapies
of brain disorders.
Standards of Practice
Assembling cohorts of cases with a brain disorder and matching
with healthy controls with respect to age, postmortem
interval (PMI), gender, hemisphere, pH and cause of death
Ruling out substance abuse with careful family history and
toxicology is essential
Learning about all the psychotropic and other medications that
each donor was taking prior to death
Determining the correct psychiatric diagnosis of the individual using
medical records and family questionnaires
Obtaining a careful neuropathological assessment to rule out the
presence of neurodegenerative disease, tumors, strokes or
trauma is essential
Table 1aS
The McLean 66 Cohort and Quality Control Parameters for RNA Integrity
Brain Number
3498
3688
3699
3706
3732
3806
3814
3817
3898
3902
3927
3933
4069
4113
4156
4157
4158
4190
4198
4237
4238
4240
4256
4337
4375
4403
4435
4462
4464
4469
4496
4545
4586
Diagnosis
Age
PMI
Schizophrenia
Control
Bipolar
Control
Control
Control
Bipolar
Bipolar
Control
Schizophrenia
Bipolar
Schizoaffective
Bipolar
Bipolar
Schizophrenia
Schizophrenia
Schizophrenia
Schizoaffective
Schizophrenia
Bipolar
Schizophrenia
Schizophrenia
Schizophrenia
Control
Schizophrenia
Bipolar
Schizoaffectiv
e
Bipolar
Bipolar
Schizophrenia
Schizophrenia
Bipolar
Control
66
66
51
40
69
70
29
64
78
83
38
84
80
42
44
35
42
78
46
78
26
42
47
36
31
76
48
50
74
80
49
74
37
22.1
18.7
31.0
28.0
15.3
15.0
10.7
11.0
14.1
23.2
41.5
25.7
12
15.8
19
28.0
18.1
13.4
18.5
30.2
16.0
27
19
24.5
29.0
22.8
33.7
30.5
24.8
10.9
19.0
7.2
18.7
Hemisphere Gender
Rt
Rt
Lt
Rt
Rt
Rt
lt
Rt
Rt
Rt
lt
Rt
Lt
lt
Lt
Lt
Lt
Lt
Lt
rt
Rt
Rt
Rt
Lt
Rt
Lt
Lt
Rt
Lt
Lt
Lt
lt
Rt
M
M
M
m
m
f
F
F
F
F
M
F
F
F
M
M
M
F
M
M
M
M
M
M
M
F
F
M
M
M
M
M
M
Brain pH
18/28s
rRNA ratio
3'/5' ratio
GAPDH
3'/5' ratio
B-Actin
Percent
Present Calls
6.43
6.76
7.02
6.50
6.88
6.59
6.70
6.69
6.22
5.91
6.52
6.14
6.51
6.26
6.05
6.25
6.26
6.81
6.31
6.30
6.75
6.64
6.57
6.26
6.46
6.60
6.63
6.35
6.53
6.44
6.60
6.70
6.68
1.80
1.50
0.95
0.55
1.50
1.35
1.30
1.30
1.26
0.25
1.45
0.50
1.00
1.30
0.94
0.96
1.00
1.20
1.04
0.70
1.60
1.20
1.90
1.14
1.08
0.53
1.28
1.05
0.80
0.50
1.80
1.30
1.35
1.00
1.13
1.8
1.7
1.3
1.18
1.06
1
1.6
1.97
1
1.67
3
1.1
3
1.63
1.32
1.5
3.2
1.9
1.3
1
2
1.5
1.21
1.9
1.19
1.6
1.4
1.4
1
2.9
1.16
1.44
1.45
3.40
3.40
2.80
2.00
1.57
1.70
2.50
3.50
1.18
2.80
4
1.70
4
2.08
2.14
2.33
2.59
4.16
2.60
2
4
2.34
2.00
3.80
1.46
4.20
3.40
1.80
1.20
4.10
1.45
48.0
43.8
43.9
46.3
43.6
46.3
45.1
50.7
42.1
42.0
43.0
39.6
27.0
52.4
34.0
46.1
46.4
46.1
45.5
42.4
49.6
45.0
47.0
44.4
49.0
42.1
47.1
46.6
47.2
41.0
50.0
33.4
49.3
PGI Case #
223
222
231
224
225
260
238
249
243
212
242
241
259
226
229
236
250
215
230
245
GENE EXPRESSION PROFILING AS A SCREENING TOOL
Quality Controls
RNAse Activity: Is the mRNA degraded?
18S:28S Ratio : A Critical Factor
Ratio  1.0  Tissue pH
Quality Control of mRNA
Schizophrenic
Control
Bipolar
17.5
30
B3933
rRNA ratio = 0.50
15.0
12.5
10.0
Triplet
#8
B3806
rRNA ratio = 1.35
25
20
B4069
rRNA ratio = 1.00
12.5
10.0
7.5
5.0
10
2.5
5
19
24
29
34
39
44
49
54
59
64
69
18S
0
28S
18S
0.0
19
Time (seconds)
24
29
34
39
0.0
28S
2.5
44
49
54
59
64
69
19
24
29
34
39
Time (seconds)
28S
5.0
18S
Fluorescence
Fluorescence
Fluorescence
15
7.5
44
49
54
59
64
69
64
69
Time (seconds)
20.0
20.0
40
B4190
rRNA ratio = 1.20
17.5
15.0
12.5
B4737
rRNA ratio = 0.80
35
30
B4545
rRNA ratio = 0.50
17.5
15.0
12.5
25
10.0
10.0
Fluorescence
Fluorescence
Fluorescence
20
7.5
7.5
15
5.0
5.0
10
2.5
2.5
19
24
29
34
39
44
49
54
59
64
0.0
69
Time (seconds)
19
24
29
34
39
44
49
54
59
64
69
19
24
29
34
39
Time (seconds)
35
B4238
rRNA ratio = 1.60
30
25
44
49
25
B4605
rRNA ratio = 1.30
20
20
59
35
B3814
rRNA ratio = 1.30
30
25
15
Fluorescence
54
Time (seconds)
20
Fluorescence
Fluorescence
15
10
10
15
5
10
34
39
44
5
49
54
59
64
0
69
0
Time (seconds)
19
24
29
34
39
44
49
54
59
64
28S
29
28S
24
18S
18S
19
28S
5
0
18S
Triplet
# 10
28S
18S
0
28S
18S
28S
5
0.0
18S
Triplet
#9
69
Time (seconds)
19
24
29
34
39
44
49
54
59
64
69
Time (seconds)
12.5
4.5
17.5
Fluorescence
Fluorescence
7.5
5.0
1.5
5.0
1.0
24
29
34
39
44
49
54
59
64
69
2.5
19
24
29
34
39
0.0
28S
18S
0.0
28S
0.5
0.0
18S
2.5
Time (seconds)
44
49
54
59
64
19
69
B4314
rRNA ratio = 0.00
5.0
4.5
4.0
3.5
29
34
39
44
54
59
64
69
64
69
15.0
B5105
rRNA ratio = 1.30
40
35
30
B4237
rRNA ratio = 0.70
12.5
10.0
25
Fluorescence
3.0
2.5
7.5
Fluorescence
Fluorescence
20
2.0
15
5.0
1.5
1.0
10
0.5
5
2.5
24
29
34
39
44
49
54
59
64
18S
0.0
28S
18S
0
19
28S
18S
0.0
69
19
Time (seconds)
24
29
34
39
44
49
54
59
64
69
19
24
29
34
39
44
Time (seconds)
7
49
54
59
Time (seconds)
17.5
4
B5083
rRNA ratio = 1.46
15.0
12.5
12.5
10.0
7.5
7.5
2
Fluorescence
Fluorescence
Fluorescence
3
5.0
5.0
1
2.5
24
29
34
39
44
49
54
59
64
69
19
24
29
34
39
Time (seconds)
0.0
28S
0.0
18S
18S
19
28S
2.5
0
B3927
rRNA ratio = 1.45
15.0
10.0
44
49
54
59
64
19
69
24
29
34
39
28S
5
18S
B4375
rRNA ratio = 1.08
6
Triplet
# 13
49
Time (seconds)
45
5.5
24
Time (seconds)
6.0
Triplet
# 12
7.5
2.0
19
B4113
rRNA ratio = 1.30
10.0
2.5
28S
10.0
3.0
Fluorescence
Triplet
# 11
3.5
18S
12.5
B4981
rRNA ratio = 1.12
4.0
B4256
rRNA ratio = 1.90
15.0
44
49
54
59
64
69
64
69
Time (seconds)
Time (seconds)
15.0
15.0
30
20
10.0
10.0
Fluorescence
Fluorescence
7.5
5.0
5.0
10
2.5
2.5
5
B4462
rRNA ratio = 1.05
12.5
7.5
15
Fluorescence
28S
18S
28S
0.0
18S
0.0
0
18S
Triplet
# 14
B4751
rRNA ratio = 1.30
12.5
19
19
24
29
34
39
44
Time (seconds)
49
54
59
64
69
19
24
29
34
39
28S
B4496
rRNA ratio = 1.80
25
44
Time (seconds)
49
54
59
64
69
24
29
34
39
44
Time (seconds)
49
54
59
Studies of Gene Expression
Using Microarrays
Are the microarray data reliable?
Replicability of Gene Expression Profiling
Comparison of Microarray Data:
1. Single Extract Run on Multiple Chips
2. Extracts From Different Lots Run at Different Times
After Freezer Storage
Data for 22,000 genes from human hippocampus!
Laser Microdissection of Hippocampal Subfields and Layers
CA4
SM = Stratum Molecular
SR = Stratum Radiatum (GABA Neurons)
SP = Stratum Pyramidale
SO = Stratum Oriens
(GABA Neurons)
Benes, Lim, Matzilevich, Walsh and Subburaju. PNAS (2007)
CA1
*
*
* = GABA Cells
SO SP SR SM
GC
Schizophrenia
Cell Cycle Regulation and
DNA Repair
Increased Expression
Increased Expression
Bipolar Disorder
Mitosis
Replication
Facto
Benes, Lim and Subburaju. PNAS (2009)
Mitosis
Functional Magnetic Resonance Imaging of Activation Networks:
Activation Networks Are Not Neural Circuits?
Amygdalar Projections to Anterior Cingulate Cortex
Van Hoesen (1993)
Van Hoesen, 1993
CA2
CA3
CA1
SUBREGIONAL CHANGES IN SZ + BD HIPPOCAMPUS
AD*
CA4
CA3
CA2**
CA1
Nonpyramidal N. (1998)
GABAA RBA (1996)
GluR5,6,7-IR
(2001)
TH-IR Varicosities (1999)
GAD65 Terminals (1998)
D1 mRNA (2004)
GAD65 +GAD67 (2007)
*
**
AD = Area Dentata
CA2 has the highest level of nuclear [3H]corticosterone binding + highest expression of
kainate receptor protein
Rodent Modeling for Postmortem Circuitry Abnormalities
in Limbic Lobe of SZs and BDs
Picrotoxin Experiments
BLn of
Amygdala
Anterogradely
Traced BLn Fibers
Str. Oriens
CA2/CA3
BLn = Basolateral Nucleus
Berretta, Munno and Benes, J. Comp. Neurol., 1999
Benes and Berretta, Neuropsychopharmacology, 2001
Excitatory Influence of Amygdala and Medial Septal Afferents
on Hippocampal Integration in SZ
Area Dentata
CA3/2
CA1
Perforant Path
Entorhinal Cx
Granule Layer
SM
Mossy Fibers
SR
Normal Control
SP
SO
Schaffer Collaterals
Alveus
Area Dentata
Schizophrenics
+
Bipolars
Septal Nuclei
Hypothalamus
CA3/2
CA1
Subiculum
Amygdala
Entorhinal Cx
GABAA Receptors
Amygdala Activation of CA3/2 in Rats
GABA
Amygdalar Glutamate
Kainate Receptors
(GluR 5 or 6)
Stratum Pyramidale
Tretrameric
Kainate Receptors
GluR 6,7
GluR 6,7
Stratum Oriens
GluR 5
Disinhibitory Interneuron
Fast-Spiking
Inhibitory Interneuron
Basolateal Amygdalar Afferents
TGFβ2
FGF2,6,9
VEGF
Neuregulin
GAD67 Network
Kainate Receptor Subunits
Cell Cycle Regulation
GRIK3
GRIK1
GRIK2
TGFβR1
LEF1
GAD67
Β-Catenin
Promoter
SMAD3
RUNX2
GSK3β
HDAC11
CDC25a
DAXX
DNA Replication/Repair
POLI
POLI
POLH
TLE1
Functional
Differentiation
Cyc E
CDK2
POLG
RPA3
Cyclin D2
CDK4/6
POLD
BRCA1
G2
p53
De-Differentiation
PAX4,5
G1 Arrest
G1
Cyc B
CDC2
E2F
Benes , FM. Cell Cycle 2010; 9: 625-627
Benes, FM. Neuropharmacology 2011; 60: 1232-42.
Abl
RB
DP-1
MBD4 CH3-CpG Domain
CHK2
OFF
No Mitosis
Oxidative Stress
Chromatin Strand
DNA Damage
DNA Polymerase
Mutations
Genomic Integrity Preserved?
Duplications or Deletions Induced?
Duplications and Deletions of Specific DNA Sequences for Important Genes
Sheng, Demers and Benes. In preparation (2011)
Informed Consent
Genomics research presents special challenges
Legal-next-of-kin (LNOK) must give consent
Irreversible anonymization provides the greatest confidentiality
BRAIN IMAGING OR POSTMORTEM STUDIES
An NIH National Resource
Which is more important for understanding psychiatric disorders?
Imaging studies tell us about function
Postmortem studies tell us about the molecular
regulation of neural circuitry
BOTH ARE EQUALLY IMPORTANT!
Office of the Inspector General
Department Health and Human Services
“There must be “trust in the tissue banking community, i.e.
organizations involved in procuring and using the donation will act as
stewards of the gift.”
HUB MODEL FOR NIH NATIONAL BRAIN BANKING
Brain Bank 2
Brain Bank 1
HBTRC
Mind Institute
UC Davis
Brain Bank
Central
Tissue Applications
Brain Bank 3
Developmental
Disorders
Maryland
Brain Bank 4
Substance Abuse
Miami