compounds known to induce robust and wide-
spread intracellular signaling in each system
( 8 ). Systemic injection of raclopride (RAC; a
D2-receptor antagonist) todrd2-eGFP mice
induced a strong nucleosomal response mostly
in D2-SPNs that extended throughout the pos-
terior striatum, whereas GBR12783 (GBR; a DA-
transporter inhibitor) induced an even stronger
activation with similar distribution but mostly
in D1-SPNs (Fig. 3A). We injected four groups
ofdrd2-eGFP mice with different combina-
tions of vehicle, RAC (t=0),andGBR(t=15)
andrecordedtheirambulatorylocomotorac-
tivity in an open field arena prior to perfusion
(t= 30) (Fig. 3B). GBR injection strongly in-
creased locomotion in both groups that re-
ceived it, irrespective of the RAC injection (Fig. 3,
C to G, and table S3). In RAC-treated mice,
taD2-SPNs dominated most of the striatal space
(Fig. 3, H and I), whereas a reverse pattern
was observed after GBR treatment (Fig. 3, H
Matamaleset al.,Science 367 , 549–555 (2020) 31 January 2020 3of7
Training days
Lever Press
Mag check
0
20
40
60
0
5
10
15
20
Training days
Cluster:
taD2-DPN
taD1-SPN
AC
B
E
F
D
GH
I
J
L
K
MNO
500 μm
AAV-FLEx-
taCasp3-TEVp
adora2a-Cre::drd2-eGFP
DARPP-32
eGFP
DARPP-32
eGFP
Merge
0 3 6 9 12 15
Mag
Extinction
CRF FR5 FR10
Instrumental
Cumulative presses Extinction
04812 20
Minutes
% of D2
Ratio D2/D1
Ratio D1/D2
% of D1
Instrumental
Extinction
16
Press/min
InstrlExt
Instrumental
Extinction
n.s.
n.s.
IAI (n+1)
IAI (n)
0.1 1 10 100
0.1
10
30
50
Press/min^10
1
Sham
100
Lesioned
IAI (n+1)
IAI (n)
0.1 1 10 100
0.1
10
1
100
03691215
Mag
Extinction
learning
CRF FR5 FR10
EC
30
50
Press/min^10
Cumulative presses
0510Min
Cumulative presses
0 5 10 15 20
Sham
LP rate Lesioned LP rate
Slp Slp
p = .1361 p < .0001
# presses
3 sec^080
(Ø)
# presses
3 sec 080
Sham Lesioned (Ø)
Extinction Learning Test
Min
Overlap
Overlap (% of D1)
taD2-SPN density
taD1-SPN density
Sham Lesioned
Sham Lesioned Sham
Lesioned
Overlap
Sham
Lesioned
DMS
Training days
0 5 10 15
0 5 10 15
n.s.
n.s.
Test
L
D
#1
#2
#3
Overlap
areas
Exclusion
Exclusion
20
40
0
20
40
0
D1-SPN
D2-SPN
D1-SPN
D2-SPN
EF-1α
TEVp taCasp3
T2A
FLEx
500 px
x
y 500 px
x
y 0
13
26
39
52
65
0
40
80
1.2
0.0
0
6
12
200
600
800
400
0 20
30
40
50
100
300
400
200
0
100
300
400
200
0
0
50
0
20
0
40
80
Instrumental
Extinction
Instrumental
Extinction
Fig. 2. Functional confluence of projection systems in the striatum
promotes extinction of learned actions.(A) Mice (eight per group) were
trained on increasing fixed ratio (FR) reinforcement schedules prior to
extinction (day 16). (B) Cumulative and average lever press performance during
instrumental (control) and extinction sessions (day 16). (C) Distribution of
taD2- and taD1-SPNs in the posterior striatum of mice in (B). Plots show up to
three density clusters from aligned hemisections in each group (4044 SPNs
mapped). (D) Reconstruction of taD2- and taD1-SPN overlapping territories in
the striatum. (EandF) Extent of taD2- and taD1-SPN territory exclusion. Data
are percentages of taD2-SPN territories segregated from taD1-SPNs (E) and
vice versa (F). Insets: overall D2/D1 (E) and D1/D2 (F) P-H3+SPN ratios.
(G) Genetic lesion of D2-SPNs in the DMS through AAV-FLEx-taCasp3-TEVp
system. DARPP-32+-eGFP-SPNs remained intact (fig. S4A). (H)Shamand
Lesioned mice were trained as in (A) but underwent additional extinction
testing on day 17. (I) Lever press performance in both groups across
instrumental training. Right: return maps of collective IAIs on day 15.
(JandK) Cumulative presses per minute on days 16 (J) and 17 (K). Insets:
average press performance (top) and linear regression slope (Slp) analysis
(bottom). (L) Raster plots and frequency histograms of pooled lever press data
preceding the delivery of each pseudoreward (red). (M) Digitized reconstruc-
tion of taD2- and taD1-SPNs after test. Insets: taD2- and taD1-SPN overlapping
territories. (N) Extent of taD2- and taD1-SPN territory overlap (% of D1).
(O)P-H3+nuclei density in D2-SPNs (left axis, purple) and D1-SPNs (right
axis, orange) in the DMS. Left: regions quantified in each group. [(E), (F), (N),
and (O)]: Two hemisections per mouse, eight mice per group; *, overall/simple
effect (black) and interaction (red). n.s., not significant (table S2).
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