Science - USA (2022-04-15)

Gabanyiet al.,Science 376 , eabj3986 (2022) 15 April 2022 2 of 12

Thalamus

Hypothalamus

AB

NeuN Iba1 Overlay

C

D

Nod2 GFP Nod2 GFP

Cortex

Striatum

NeuN Iba1 Overlay

CPu

MGN

LGN

Po

CPu

Acc

OT

CPu

Acc

SN

OT

VM

Hyp

OT POA

VTA

IC

% of

double positiv

e NeuN

NeuN+GFP+ +

Iba1+GFP+Iba1+

E F

0

50

100

150

200

250 Brain

*

0

50

100

150

Counts per minute

Blood

E. coli

FB8-lysA

gavage

24h

Wash

4x

(^3) H mesoDAP
H 2 O + Streptomycin
Ctr
Cortex Striatum Thalamus Hypothalamus
/
/
0
10
20
30
40
80
100
Ctr
24h
GFP (Nod2) GFP (Nod2)
Cortex
Hypothalamus
Striatum
Thalamus
Fig. 1. Nod2 and Nod2 ligands in the brain.(AandB) Immunofluorescence
ofNod2GFPmouse brain slices, as shown in representative images of brain sagittal
slices stained for Nod2 (GFP, white) (A) and a coronal slice stained for neurons
(NeuN, magenta), microglia (Iba1, white), and Nod2 (GFP, green) (B), highlighting
cortex, thalamus, striatum, and hypothalamus. Scale bars, 150mm. (C) Quantification
of neurons (NeuN+), microglia (Iba1+), and their colocalization with Nod2 (GFP+)
in the cortex, striatum, thalamus, and hypothalamus. (D) Colocalization of Nod2
(yellow) with Iba1 (cyan) or NeuN (magenta) in the cortex, thalamus, striatum, and
hypothalamus. Pink arrowheads indicate Iba1+Nod2+cells; white arrowheads indicate
NeuN+Nod2+cells. Scale bars, 10mm. (E) PG tracking experiment scheme.
(F) Radioactivity detected 24 hours after gavage with^3 H-labeledE. coliin the brain
and blood of older females and males (7 to 8 months;n= 6 per group). Controls
were gavaged with nonlabeledE. coli(n=4).Dataareaverages±SEM.*P≤0.05
(unpairedttest). Abbreviations: CPu, caudate putamen; Hyp, hypothalamus; IC,
inferior colliculus; LGN, lateral geniculate nucleus; MGN, medial geniculate nucleus;
Acc, nucleus accumbens; OT, olfactory tubercles; Po, posterior thalamic nucleus; POA,
preoptic area; SN, substantia nigra; VM, ventral medial nucleus of the thalamus;
VTA, ventral tegmental area; PG, peptidoglycan.
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