Science - 31 January 2020

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

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Training days

Cluster:

taD2-DPN

taD1-SPN

AC

B

E

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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

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Extinction

16

Press/min

InstrlExt

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Extinction

n.s.

n.s.

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IAI (n)

0.1 1 10 100

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Press/min^10

1

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IAI (n)

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03691215

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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

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(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

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areas

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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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