1Confidential
Shank 3 Mouse Model of Autism
2Confidential
Background
• About 2% of people with autism carry harmful mutations in SHANK3, a protein that helps organize the connections between neurons
• Developed by Guoping Feng, Shank3 –Feng mice harbor a deletion of exons 13-16 of the PDZ domains leading to the deletion of the Shank3α and Shank3β isoforms and partial deletion of Shank3γ
• These mice exhibit some social, communicative, repetitive, and sensory processing abnormalities associated with autism spectrum disorder.
• Displays some social deficits• Shows a decrease in activity and locomotion compared to littermate controls,
including reduced rearing• Shows gait differences compared to littermate controls• Displays some anxiety-like behaviors• Shows reduced startle responses and increased prepulse inhibition of startle
3Confidential
No Significant differences in Body Weight Between WT and KO Mice
Age (weeks)
5 6 7 8 9 10 11
Bo
dy W
eig
ht
(g)
10
15
20
25
30
SHNAK3/F_WTSHNAK3/F_KO
4Confidential
Behavioral Assessments
5Confidential
KO Mice Show Deficits in Rotarod PerformanceL
ate
nc
y t
o F
all
(s
ec
)
0
50
100
150
200
Shank3/F_WTShank3/F_KO
*
Sp
eed
at
Fall
(rp
m)
0
10
20
30
Shank3/F_WTShank3/F_KO
*
6Confidential
Urine Open Field Test
• Male mice are placed in an Open Field where they are presented with the urine of an estrus female
• Activity, proximity to the urine, ultrasonic vocalizations, and urine marking by the male are recorded to determine social and function
• Urine spots developed with Nindyrin Spary
Estrous female urine spot
7Confidential
KO Mice Show Decreased Activity in the Open Field Following Exposure to Female Urine
To
tal
Dis
tan
ce T
ravele
d (
cm
)
0
300
600
900
1200
1500
SHNK3/F_WTSHNK3/F_KO
*
Dis
tan
ce
Tra
ve
led
in
Ce
nte
r (c
m)
0
100
200
300
400
500
SHNK3/F_WTSHNK3/F_KO
*%
Tim
e S
pen
t in
Ce
nte
r
0
10
20
30
SHNK3/F_WTSHNK3/F_KO
*
8Confidential
Reciprocal Social Interaction
• Same-genotype, -sex, -age dyads are allowed to freely interact for 10 minutes
• Distance between mice, proximity, and interactions are measures of sociality
Front Back Side
Nu
mb
er
of
Inte
racti
on
s
0
10
20
30
40
50
Shank3/F_WTShank3/F_KO
* *
Tim
e S
pe
nt
Fo
llo
win
g (
sec)
0
10
20
30
40
50
60
70
80
Shank3/F_WTShank3/F_KO
*
9Confidential
KO Mice Show Increased Grooming
Gro
om
ing
Bo
uts
0
30
60
90
120
150
Shank3/F_WTShank3/F_KO
*
10Confidential
KO Mice Show Decreased Startle Response
Me
an
Sta
rtle
Re
sp
on
se
0
200
400
600
800
1000
1200
Shank3/F_WTShank3/F_KO
*
11Confidential
KO Mice Show Increased Hindlimb Clasping
Mice are lifted gently by the tail with front limbs just above the surface
Clasping of hind legs is noted (normal is a spread in the hind legs)
Pe
rcen
t M
ice
Cla
sp
ing
0
10
20
30
40
50
60
70
80
90
100
Shank3/F_WTShank3/F_KO
*
12Confidential
KO Mice Show Decreased Ultrasonic Vocalizations
Ult
ras
on
ic V
oca
liza
tio
ns
0
30
60
90
120
150
Shank3/F_WTShank3/F_KO
*
13Confidential
Electrophysiology
14Confidential
Impaired corticostriatal synaptic transmission in Shank3 KO
• Extracellular field potential responses (A) and whole-cell currents in medium spiny neurons (B) were evoked in dorsal striatum by
stimulating corpus callosum.
• Brain slices were prepared from male mice aged 10 weeks for extracellular field potential recordings (A) and 14 weeks for whole-
cell patch clamp recordings (B).
• Numbers in parenthesis show number of slices (cells); animals used.
A B
15Confidential
Impaired NMDA-mediated synaptic transmission in dorsal striatum in Shank3 KO
• Whole-cell NMDA-mediated currents in medium spiny neurons were evoked in dorsal striatum by stimulating corpus
callosum in 14 weeks old male mice.
• Numbers in parenthesis show number of cells; animals used.
16Confidential
Impaired synaptic transmission in nucleus accumbens in Shank3 KO
• Extracellular field potential responses were evoked in nucleus accumbens (NAc) by stimulating terminals within the
nucleus in 10 weeks old male mice.
NAcacp
Str
17Confidential
Molecular markers
18Confidential
Decreased expression of synaptic proteins in striatum and BNDF in cortex in Shank3 KO
• Expression of synaptic proteins in striatum and BDNF in cortex was assessed by qPCR in 16 weeks old male
Shank3 KO mice (n=10).
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