Science - USA (2022-04-15)

We then performed a 3-day differential au-
ditory FC paradigm in a separate group of
mice while recording somatic and dendritic
Ca2+activity in head-fixed mice under a two-
photon microscope (Fig. 4A). Animals exhibited
increased levels of freezing behavior when

exposed to the CS 24 hours after FC under

freely moving conditions before imaging Ca2+

responses (Fig. 4B). In these animals (n=9),

we imaged in total 177 neurons, 82% of which

showed CS responses before or after condi-

tioning, whereas 18% of all neurons were not

CS responsive (fig. S6C). Compared with pre-

conditioning levels, 84% of all CS responsive

neurons (123 neurons from nine animals) ex-

hibited learning-associated plasticity of CS

responses, whereas 13% showed stable CS re-

sponses (fig. S6C).

dÕAquinet al.,Science 376 , eabf7052 (2022) 15 April 2022 4 of 13

Soma

Dendrite

2 z-score

30 s

Saline

CNO

Soma

Dendrite

Day 1:

Day 2:

Saline

CNO

GRIN lens

LA

SST-Cre

0.2

0.4

0.6

0.8

1

0

Soma-dendrite transients amplitude correlaltion (r)

***

SalineCNO

hM4D(Gi)-mCherry

50 μm

GCaMP6s

2

4

6

8

0

Dendrite only

transients per min

***

SalineCNO

10

4

8

0

12

Transients

per min

n.s

SalineCNO

16

hM4D

GCaMP hM3D GCaMP

mCherry

GCaMP

(^25) 048 048
Soma #
-1
0
1
F/F
Time (s) from CS onset
Habituation Conditioning
2 s
10%
F/F
**
0
0.4
0.8
0
0.4
0.8
1.2
Hab.Cond. Hab.Cond.
CS response
integral (
F/F
)
CS response
amplitude (
F/F
) *
Habituation
Conditioning (CNO)
(^12) 048 048
Dendrite #
-1
0
1
F/F
Time (s) from CS onset
Habituation Conditioning
0.4
0.8
1.2
0
Hab.Cond.
CS response
integral (
F/F
)
n.s
0.2
0.4
0.6
0
Hab.Cond.
CS response
amplitude (
F/F
)
n.s
2 s
10%
F/F
Habituation
Conditioning (CNO)
(^26) 048 048
Soma #
-1
0
1
F/F
Time (s) from CS onset
Habituation Conditioning
0
1
0.4
0.8
2
0
Hab.Cond. Hab.Cond.
CS response
integral (
F/F
)
CS response
amplitude (
F/F
)
n.s n.s
2 s
10%
F/F
Habituation
Conditioning (CNO)
Habituation
Conditioning (CNO)
2 s
10%
F/F 0.5
1
1.5
0
2
Hab.Cond.
CS response
amplitude (
F/F
)

0
1
2
3
Hab.Cond.
CS response
integral (
F/F
)
**
048
Time (s) from CS onset
Habituation Conditioning
048
Dendrite #
-1
0
1
F/F
25

---

%
Dendrite-only
transients
20
40
60
80
100
0
Spontaneoustone-locked
CNO
Saline
n.s
0.2
0.6
0.4
0.8
0
SalineCNO
1
0.2
0
n.s
Transients amplitude Transients integral
(norm.
F/F)
(norm.
F/F)
0

• SalineCNO
  0
  0.2
  0.4
  0.6
  E
  A B C
  F
  D
  HIJ
  GP
  N O
  Q R
  L M
  S T
  K
  PN PN
  PN
  SST+ SST+
  SST+
  Fig. 3. SST+ interneurons control dendritic activity and CS responses.
  (A) Viral approach to express hM4D(Gi) in SST+ INs and GCaMP6s in a sparse
  population of LA PNs. Ninety tones were presented after saline or intraperitoneal
  injection of CNO on 2 consecutive days. (B) Experiment scheme. (C) Confocal
  image of hM4D(Gi)-mCherry in SST+ INs with sparse GCaMP6s in LA PNs.
  (D) Reduction of SST+ IN output increases dendrite-only transients. Arrow-
  heads indicate compartment-specific transients in an example soma-dendrite
  pair. (E) Reduction of SST+ IN output increases the rate of dendrite-only transients
  (P< 0.001 Wilcoxon signed-rank test,n=79 dendritic segments, 14 neurons).
  (F) Reduction of SST+ IN output increases spontaneous, but not tone-locked,
  dendrite-only transients (Spontaneous,P< 0.001; tone-locked,P= 0.125,
  Wilcoxon signed-rank test). (G) Reduction of SST+ IN output reduces the
  amplitude correlation between somatic and dendritic transients (r, Pearson’s
  correlation coefficient,P< 0.001 Wilcoxon signed-rank test). (H) Reduction of
  SST+ IN output does not change the rate of somatic transients (P= 0.347,
  Wilcoxon signed-rank test,n=30 neurons). (I) Reduction of SST+ IN output does
  not change the amplitude of somatic transients (P= 0.140, Student’sttest).
  (J) Reduction of SST+ IN output decreases the integral of somatic transients
  (P= 0.022, Student’sttest). (K) Experimental scheme. (L) (Left) CS responses
  ordered according to amplitude during habituation. (Right) Mean responses ±
  SEM (n=25 CS-responsive dendrites during habituation, five mice). (M) Statistical
  analysis of (L). (Left) CS responseDF/Famplitude (P= 0.002 Wilcoxon matched-pairs
  signed-rank test). (Right) CS responseDF/Fintegral (P= 0.705 Wilcoxon matched-
  pairs signed-rank test; conditioning, 0.307 ± 0.096DF/Fintegral). (N) (Left) CS
  responses ordered according to amplitude during habituation. (Right) Mean
  responses ± SEM (n=25 CS-responsive somas during habituation, 5 mice).
  (O) Statistical analysis of (N). (Left) CS responseDF/Famplitude (P< 0.001 Wilcoxon
  matched-pairs signed-rank test). (Right) CS responseDF/Fintegral (P= 0.002
  Wilcoxon matched-pairs signed-rank test). (P) Experimental scheme. (Q) (Left)
  CS responses ordered according to amplitude during habituation. (Right) Mean
  responses ± SEM (n=12 CS-responsive dendrites during habituation, four mice).
  (R) Statistical analysis of (Q). (Left) CS responseDF/Famplitude (P= 0.979
  Student’sttest). (Right) CS responseDF/Fintegral (P= 0.733 Wilcoxon matched-
  pairs signed-rank test). (S) (Left) CS responses ordered according to amplitude
  during habituation. (Right) Mean responses ± SEM (n=26 CS-responsive somas
  during habituation, four mice). (T) Statistical analysis of (S). (Left) CS response
  DF/Famplitude (P= 0.585 Student’sttest). (Right) CS responseDF/F integral
  (P= 0.568 Wilcoxon matched-pairs signed-rank test).
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