|
Display additional annotations [+]
Gene |
Clone |
Species |
Stages |
Anatomy |
irx3
|
|
laevis
|
NF stage 15
|
chordal neural plate
,
preplacodal ectoderm
|
irx3
|
|
laevis
|
NF stage 20
|
posterior neural tube
,
anterior placodal area
|
irx3
|
|
laevis
|
NF stage 28
to
NF stage 35 and 36
|
brain
,
midbrain
,
hindbrain
|
barhl2
|
|
laevis
|
NF stage 15
|
neural plate
|
barhl2
|
|
laevis
|
NF stage 20
|
neural tube
|
barhl2
|
|
laevis
|
NF stage 28
to
NF stage 35 and 36
|
retina
,
forebrain
,
eye
|
arx
|
|
laevis
|
NF stage 15
|
neural plate
|
arx
|
|
laevis
|
NF stage 20
|
neural tube
|
arx
|
|
laevis
|
NF stage 28
to
NF stage 35 and 36
|
forebrain
|
pax6
|
|
laevis
|
NF stage 15
|
chordal neural plate
,
anterior neural fold
|
pax6
|
|
laevis
|
NF stage 20
|
optic vesicle
,
posterior neural tube
,
neural tube
|
pax6
|
|
laevis
|
NF stage 28
to
NF stage 35 and 36
|
brain
,
forebrain
,
eye
|
lhx2
|
|
laevis
|
NF stage 15
|
anterior neural fold
|
lhx2
|
|
laevis
|
NF stage 20
|
optic vesicle
|
lhx2
|
|
laevis
|
NF stage 28
to
NF stage 35 and 36
|
retina
,
brain
,
forebrain
,
midbrain
,
hindbrain
,
[+]
|
lhx9
|
|
laevis
|
NF stage 20
|
optic vesicle
,
anterior neural tube
,
neural tube
|
lhx9
|
|
laevis
|
NF stage 28
to
NF stage 35 and 36
|
retina
,
brain
,
forebrain
,
midbrain
,
hindbrain
,
[+]
|
rax
|
|
laevis
|
NF stage 15
|
anterior neural fold
|
rax
|
|
laevis
|
NF stage 20
|
optic vesicle
|
rax
|
|
laevis
|
NF stage 28
to
NF stage 35 and 36
|
retina
,
eye
|
fezf2
|
|
laevis
|
NF stage 15
|
anterior neural fold
|
fezf2
|
|
laevis
|
NF stage 20
|
olfactory placode
|
fezf2
|
|
laevis
|
NF stage 28
to
NF stage 33 and 34
|
forebrain
|
|
|
Figure S2. Comparison of expression pattern of fezf2 expression relative to various
nerual markers at different stages, related to Figure 1. Anterior is to the bottom in
stage 15 and 20 embryos (frontal view), and to the left in stage 28 and 35 embryos
(lateral view). |
|
Display additional annotations [+]
|
|
Figure S4 (related to Figure 1). fezf2 knockdown, but not control MO knockdown,
leads to defects in forebrain neuronal differentiation. (A) fezf2 knockdown led to
defects in forebrain development after initial patterning stage. Whole-mount in situ hybridisation analyses on control versus fezf2 MO injected stage 28 embryos, stained
using the forebrain-specific markers, otx2 and pax6. (B) fezf2 knockdown did not
cause defects in early forebrain patterning, as revealed by whole-mount in situ
hybridisation on control versus fezf2 MO injected stage 17 embryos, stained using the
forebrain-specific markers arx, otx2, and pax6. Anterior is to the bottom. (C-H) 1 of 2
blastomeres at 2-cell stage was injected with control MO, cultured to stage 30, fixed
sectioned transversely across the forebrain and stained via immunofluorescence for
the transcription factor Sox3 (marker for neural progenitors) (C-D) or MyT1 (marker
for differentiated primary neurons) (F-G). (C) Merged image. (D) Sox3 staining alone.
(E). Statistical analysis of Sox3+ cells in control MO injected side relative to the
uninjected side of the forebrain (n=3 embryos, no significant difference). The
uninjected side has been normalised to 100%. (F) Merged image. (G) MyT1 staining
alone. (H) Statistical analysis of MyT1+ cells in control MO injected side relative to
the uninjected side of the forebrain (n=3 embryos, no significant difference). The
uninjected side has been normalised to 100%. (I-K) Transverse sections were
prepared as for panels C-D and then processed for TUNEL staining. (I) Merged image.
(J) TUNEL-positive cells alone. (K) Percentage of the TUNEL-positive cells in the
control MO injected side compared to the uninjected side (n=3 embryos). In all cases,
the FITC tag on the control MO was used to identify the injected side; DAPI was used
to stain nuclei. Error bars represent ±s. e. m. Scale bar: 25 μM. ns: not significant. (L)
(a-b) Immunofluorescence staining using the neuronal differentiation marker, Ntubulin,
on stage 32 embryos injected with 10 ng of control or fezf2 MO. (a) control
MO; (b) fezf2 MO, showing a strong reduction of N-tubulin staining in the rostral area.
(c) Sections of stage 32 embryos at the prethalamus level and stained with N-tubulin
antibody. Embryos were injected with 5 ng of fezf2 MO into one of the two blastomeres at the 2-cell stage. A strong reduction of N-tubulin staining was observed
in the injected side. Arrowhead: injected side. DAPI was used to provide global
visualisation on the head structure. In all cases, error bars represent ±s. e. m. |