Science - USA (2019-01-04)

silencing with that of V1 silencing, we also pre-
sented drifting gratings (Fig. 3, E and F) iden-
tical to those used to measure V1 silencing (Fig. 1,
C and D). Optogenetic silencing of SC suppressed
the response of POR to drifting gratings (Fig. 3F),

highlighting the much stronger impact of SC than

V1 on visually evoked activity in POR (79.83 ±

4.46% average decrease ± SEM in visually evoked

firing rate;P< 0.0001, Wilcoxon signed-rank test,

n= 44, 2 mice).

Because SC sends sparse inhibitory GABAergic

projections to the dLGN ( 21 ), our optogenetic

activation of GABAergic projection neurons in

SC might suppress dLGN neurons. The dLGN

projects to other visual areas in addition to V1 ( 16 )

Beltramoet al.,Science 363 ,64–69 (2019) 4 January 2019 3of6

Fig. 2. Anatomical and
functional segregation
of cortical and collicular
inputs in the pulvinar.
(A) (Top) Coronal
sections showing the
injection sites of AAV1.
CAG.TdTomato in V1
and AAV1.CAG.GFP in
SC (left) and of V1 and
SC projections to
dLGN and pulvinar
(Pulv) for three repre-
sentative sections
along the rostro-caudal
axis. (Bottom left)
Normalized fluorescent
density (average ± SEM)
along rostro-caudal axis
of the pulvinar (green,
SC input; magenta,
V1 input; 54 coronal
sections, three mice).
Fluorescence density
values were normalized
to those for the coronal
section displaying
maximal tdTomato or
GFP expression. (Bottom
right) Schematic
illustration of the results.
(B)(Toprow)The
stimulus was a dark dot
moving along a straight
trajectory. Recordings
were performed simulta-
neously from stratum
opticum of SC (with an
optrode for silencing) and
caudal pulvinar. (Second
row) (Left) PSTH of
an SC unit. Black, control;
blue, SC silencing; black
horizontal bar, period
of stimulus presentation;
blue horizontal bar, period
of SC silencing. (Right)
PSTH of a simultaneously
recorded unit in the
caudal pulvinar. (Third
row) Average PSTH
for 33 (left) and 34 (right)
isolated units in SC and
in caudal pulvinar (five
animals). (Bottom row)
Scatter plots of firing
rates averaged over a 450-ms interval (i.e., the response window; see
materials and methods for details on the moving dot analysis) during the
period of visual stimulation. Green data points, example units shown in the
second row. (C) (Top row) V1 silencing and simultaneous recordings in

caudal pulvinar. (Second row) PSTH of a unit in caudal pulvinar. Black,

control; blue, V1 silencing. (Third row) Average PSTH for 63 isolated units

(seven animals). (Bottom row) Scatter plot as in (B). (D) Similar experiment

as shown in (C) but recording from rostral pulvinar (16 units; three animals).

Caudal Rostral

Rec. Pulv

LED

V1

Laser Rec.

Rec.

SC

Caudal Rostral

Pulv

V1

Caudal Rostral

Pulv

Rec.

LED

V1

B

dLGN

500 μm

Bregma: -2.52mm

dLGN

500 μm

Bregma: -2.04mm

Caudal Rostral

SC

0102030

0

10

20

0 5 10 15 20

Pulv

500 μm

Bregma: -3.12mm

Normalized Fluorescence Density

Caudal Rostral

1

0.8

0.6

0.4

0.2

(^0) -3.2 -2.8
-2.4 -2.0
Distance from bregma (mm)
SC to pulvinar
V1 to pulvinar
C.Pulv.firing rate-LED V1 ON (Hz)
C.Pulv..firing rate-LED V1 OFF (Hz) R.Pulv..firing rate-LED V1 OFF (Hz)
0
5
10
15
A
Firing rate (Hz)
30 20
0
5
10
15
0 5 10 15 20
C.Pulv..firing rate-laser SC OFF (Hz)
20
0 5 10 15
0
5
10
15
0 +1 +2
0
5
10
15
0
5
10
15
Firing rate (Hz)
0
0.2
0.3
0.1
0.4
0.5
0.6
0
0.2
0.3
0.1
0.4
0.5
Normalized firing rate
0.6
500 μm
V1 SC
a
Bregma: -3.9mm
V1: AAV1-CAG-TdTomato
SC firing rate-laser SC ON (Hz)
C.Pulv.firing rate-laser SC ON (Hz) R.Pulv.firing rate-LED V1 ON (Hz)
SC firing rate-laser SC OFF (Hz)
SC
(stratum opticum)
Caudal Pulv
(-3.2mm bregma -2.4mm)
CD
SC: AAV1-CAG-GFP
Pulv
Pulv
Injection sites
Silencing SC Silencing V1 Silencing V1
single unit single unit single unit single unit
average average average average
SC
Pulv
Caudal Rostral
+3
Time from stimulus onset (s)
0 +1 +2 +3
Time from stimulus onset (s)
0 +1 +2 +3
Time from stimulus onset (s)
0 +1 +2 +3
Time from stimulus onset (s)
Caudal Pulv
(-3.2mm bregma -2.4mm)
Rostral Pulv
(-2.3mm bregma -1.8mm)
Normalized firing rate Normalized firing rate Normalized firing rate
Firing rate (Hz) Firing rate (Hz)
RESEARCH | REPORT
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