Science - USA (2022-01-28)

Paredeset al.,Science 375 , eabk2346 (2022) 28 January 2022 7 of 10

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i12b-GFP

time lapse imaging

NeuN SST HNA Merge

NeuN PV HNA Merge

10

A B

(^04590)
2
4
6
8
10
Total Number of DCX+ Processes
DAT
DCX+ Processes

(^04590)
500
1000
1500
Total Length of DCX+ Processes
DAT
Microns (
μm

(^04590)
5
10
15
20
Nodes on DCX+ Processes
DAT
Nodes per DCX+ Process

45 DAT 90 DAT
HNA
+/DCX+
C
F NeuN/CB/HNA
5 NeuN/CR/HNA
(^63)
D AT
G
(^56)
3
D AT
0-1011-2021-3031-4041-5051-6061-7071-8081-90
0
10
20
30
40
Speed (μ
Total GFP+ Cells (%)
0-^505 1-100101-^150 151-^2 201-^0022550 1-300 301 - 351-400^350401 -^45450 1-500501-^550 551-600601-650
0
10
20
30
40
Total Distance Traveled (μm)
Total GFP+ Cells (%)
GABA DCX HNA Merge
5
5
m
D
HNA DCX Merge
E
T
A
D
0
9
T
A
D
(^09)
Fig. 5. Transplanted hMGE cells display migratory and functional inter-
neuronal features.(A) Neurolucida tracings of DCX+transplanted hMGE at
45 and 90 DAT (four cells shown per time point). (B) Quantification of the
length (left) and total number (center) of DCX-positive processes at 45 and
90 DAT. Right graph shows quantification of the number of DCX+branching
nodes per transplanted cell analyzed at 45 or 90 DAT. For all graphs,n=
22 cells per animal for both 45 and 90 DAT; ****P< 0.0001. (C) Left:
Sequential images of time-lapse confocal microscopy showing two
example sets (red and yellow arrows) of GFP+transplanted hMGE cells at
21 DAT. Right: Proportion of GFP-positive transplanted hMGE cells (total
395 cells) analyzed for speed (left) and total distance traveled (right).
(D) Confocal image of transplanted mouse cortex at 90 DAT showing
dispersal of HNA+DCX+cells. Inset shows boxed area at higher magnification.
(E) Top: Injection site at 90 DAT contains cells that coexpress DCX, GABA,
and HNA. Bottom: Transplanted hMGE cells away from the injection site
also coexpress DCX and GABA (arrows). (F) Examples of transplanted
hMGE cells at 365 DAT that express NeuN (green) and HNA (blue) either
with SST (red, arrowheads in top panel) or PV (red, arrowhead in bottom
panel). (G) Examples of transplanted hMGE cells at 365 DAT that
express NeuN, calbindin (CB+, arrowhead in top), and calretinin (CR+,
arrowhead in bottom). Scale bars, 50mm [(A), (C) (leftmost panel), (E)],
25 mm [(C) (upper and lower rows)], 10mm [(D) (inset), (F), (G)].
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