Science - USA (2022-01-07)

and stimulus-evoked activity during altered sen-
sory experience using transgenic fosGFP mice
( 30 ) along with virally coexpressed Rcamp1.07
in S1 (n=3)(Fig.3,BandC).ExvivoHCR-FISH
confirmedthathighfosGFPfluorescencecor-
responded with higherFosmRNA and that
green fluorescent protein (GFP) mRNA was
also enriched in Baz1a neurons (Fig. 3D and
fig. S18A). Although estimated event rates

correlated with FosGFP expression levels in

Adamts2 and Agmat neurons, they did not in

Ba1za neurons, confirming thatFosdoes not

necessarily reflect ongoing activity in this cell

type (fig. S18B).

FosGFP and sensory-evoked calcium re-

sponses were tracked before and for 5 days

after bilateral whisker deprivation (BD). During

BD, the principal whisker corresponding to

the imaged S1 barrel column was trimmed to

a minimum length so that stimulus-evoked

activity could still be tracked (fig. S17) ( 31 ).

Overall, BD resulted in a decrease in stimulus-

evoked activity after 1 day followed by a slow

homeostatic compensation after 5 days [P<

0.0002, one-way analysis of variance (ANOVA),

post hoc multiple comparison test] (Fig. 3E)

( 32 ). We first asked how fosGFP expression

Condyliset al.,Science 375 , eabl5981 (2022) 7 January 2022 4of9

Fig. 3. Persistent IEG expression and homeostatic
plasticity in Baz1a neurons.(A) Examples of
selectively (top) enriched and (bottom) not enriched
immediate early genes in Baz1a cells. (B) Time
course of bilateral whisker deprivation (BD) experiment.
(C) (Top) Example of Baz1a neuron with stable high
fosGFP expression across in vivo imaging sessions.
(Middle) Average stimulus responses during calcium
imaging. (Bottom) Post hoc identification of neuron
and HCR-FISH for select genes. (D) HCR-FISH
Fosspot density in high (1.2-fold above background)
and low fluorescent fosGFP cells (two-tailed Student’s
ttest). (E) Mean stimulus-evoked activity before
and after BD across functionally imaged neurons
(one-way ANOVA with post hoc multiple comparison
test,n= 2569 cells from three animals). (F) Change
in stimulus-evoked responses before BD versus at
1 day or 5 days BD across neurons with stable low,
dynamic, and stable high fosGFP expression (two-tailed
Student’sttest,n= 790 cells from three animals).
(G) Change in stimulus-evoked responses before BD
versus at 1 day or 5 days BD across excitatory cell types
(c^2 test,n= 181 Adamts2, 136 Baz1a, and 153 Agmat
cells from three animals). (H) Fraction of fosGFP
neurons with stable high expression across all pre-BD
sessions (days–2,–1, and 0) and across all BD sessions
(days 1, 3, and 5) for excitatory cell types (two-tailed
Student’sttest,n= 3753 cells from three animals).
*P< 0.05, **P<0.005 in (E) to (G). Error bars = SEM;
(H) SD from bootstrap analysis.

25

0

Stable high

(% fosGFP cells)

Est. event rate (Hz)

0.10

0.07

BD

Session (day)

-2 -1 0 1 5

Adamts2 Baz1a Agmat

Pre-BD BD

1d BD 5d BD

Stable low Dynamic Stable high

Fos mRNA (x10-³ spots/μm^3 )

cpd.

1

0

08

A

BC

D

E

H

F

G

1d BD 5d BD

High fosGFP

Low fosGFP

Adamts2 Baz1a Agmat

Cell types

fosGFP

Δ Stim. response

Δ Stim. response

1

0

-1

1

0

-1

Days

fosGFP

Rcamp1.07

BD

-2-1 0 1 3 5

0.5

Hz

2 s

Adamts2 Baz1a Agmat

Enriched IEGs

Not enriched IEGs

Fosb Fosl2 Nr4a3 Egr2 Nr4a2 Nptx2 Fos

Cebpb Arc Scg2 Nefm Vgf Syt4 Jun

10

0

log

10

CPM

10

0

log

10

CPM

pre-BD

**

**

*

*

**

**

**

**

*

*

RCamp1.07

fosGFP

fosGFP

Rcamp1.07

Activity

-2 -1 0 1 3 5

In vivo

Ex vivo

Coch

S100a6

Rrad

Fos

Ngb

Fst

Penk

Days

P < 1x10-19

RESEARCH | RESEARCH ARTICLE