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Fig. 6. Immunohistochemistry of necklace and ectopic glomeruli. (A)
Sections (18 µm) through the caudal OB at the level of the necklace area of
a GCD-ITG homozygous mouse stained with an antibody to PDE2A. All
GFP+ axons (green) are also PDE2A+ (red), and are thus
yellow in the overlay image. Right-most panel: large glomerulus. Dorsal is up.
(B) Sections (18 µm) through the caudal OB at the level of the
necklace area of a GCD-ITL homozygous, NP2-
heterozygous mouse stained
with an antibody to ß-galactosidase (red). A subset of Nrp2+
(GFP; green) glomeruli is also positive for ß-galactosidase (arrows);
other necklace glomeruli are only Nrp2+ (asterisks). All
GC-D+ glomeruli are also Nrp2+ (yellow in overlay).
Right-most panel: high magnification. Dorsal is up. (C) Sections (18
µm) through the OB of a GCD-ITG homozygous, F homozygous mouse
stained with antibodies to OMP (red). Green is the intrinsic green
fluorescence of GFP. Three left panels: a high-magnification view of an
ectopic glomerulus that is filled by GC-D+ axons (yellow in
overlay), at the midlevel of the OB outside the necklace area. Right-most
panel: example of glomeruli in the necklace area. A GC-D+
glomerulus (red and green) above a GC-D- glomerulus (red only).
Ectopic glomeruli reside deeper than glomeruli in the necklace area. Dorsal is
up. (D) Sections (18 µm) through the midlevel OB outside the
necklace area of a GCD-ITL homozygous, NP2- homozygous (GFP; green)
mouse stained with antibodies to MAP2 (red) and ß-galactosidase (blue).
MAP2 stained dendrites fill the glomeruli in the glomerular layer (GL)
including the ectopic GC-D+/Nrp2+ glomerulus (light blue
in overlay image), but are absent in the outer nerve layer (ONL). Scale bars:
100 µm in A,B,D; 50 µm in C.
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