|
Display additional annotations [+]
Gene |
Clone |
Species |
Stages |
Anatomy |
dll1
|
|
laevis
|
NF stage 17
|
neural plate
,
anterior neural fold
,
otic region
|
eya1
|
|
laevis
|
NF stage 17
|
pre-chordal neural plate
,
anterior neural fold
|
six1
|
|
laevis
|
NF stage 17
|
pre-chordal neural plate
,
anterior
|
sox3
|
|
laevis
|
NF stage 17
|
neuroectoderm
,
trigeminal placode
,
neural plate
|
neurog2
|
|
laevis
|
NF stage 17
|
chordal neural plate
,
trigeminal placode
,
neuron
,
otic region
|
tubb2b
|
|
laevis
|
NF stage 17
|
chordal neural plate
,
trigeminal placode
,
neuron
|
neurod1
|
|
laevis
|
NF stage 17
|
chordal neural plate
,
trigeminal placode
,
neuron
|
neurod1
|
|
laevis
|
NF stage 28
|
olfactory placode
,
otic vesicle
,
optic vesicle
,
epibranchial placode
,
lateral line placode
|
cdknx
|
|
laevis
|
NF stage 17
|
chordal neural plate
,
anterior neural fold
,
otic region
|
|
|
Fig. 2. Effects of Eya1 and Six1 knockdown on markers of neurogenesis and placodal ectoderm. (A) Immunoblot showing that Eya1MO1 but not control MO blocks synthesis of Eya1 protein. (B) Co-injecting Eya1MO2 or Eya1MO1+2 with myc-tagged Eya1 mRNA significantly restores NeuroD and prevents ectopic p27Xic1 expression (χ2 test; *: p < 0.05, **: p < 0.001) in three independent rescue experiments. (C–Q) Neural plate stage embryos after unilateral injection (lower half) of Eya1MO1+2 (C–L) or Six1MO1+2 (M–Q). In all cases, where myc-GFP was co-injected as lineage tracer (C–K), embryos are shown superimposed with green fluorescent channel. Asterisks indicate reductions, arrows and arrowheads indicate increased marker gene expression. For Delta1 after Eya1MO1+2 injections two different phenotypes are depicted (J, K). R: Tail bud stage embryo after unilateral injection (R1, R3, R5: injected side; R2, R4, R6: control side) of Eya1MO1+2 reveals reduction of NeuroD expression in all neurogenic placodes or derivative ganglia. R3 and R4 depict a section at the level indicated (white line) with boxed areas shown magnified in R5 and R6, respectively, superimposed with green (myc-GFP co-injected with Eya1MO1+2) and blue (DAPI) fluorescence. Residual NeuroD expression is confined to cells receiving little or no MO. Bar in R4: 100 μm (for R3, R4). Abbreviations: e: eye; epVII+AV: facial epibranchial and anteroventral lateral line placode; epIX: glossopharyngeal epibranchial placode; gVII/AV/AD: ganglion of the facial, anteroventral and anterodorsal lateral line nerve; gVIII: vestibulocochlear ganglion; nt: neural tube; pAD: anterodorsal lateral line placode; pM: middle lateral line placode; pOl: olfactory placode; pP: posterior lateral line placode; pPr: profundal placode; pV: trigeminal placode; vOt: otic vesicle. |
|
Display additional annotations [+]
Gene |
Clone |
Species |
Stages |
Anatomy |
dll1
|
|
laevis
|
NF stage 17
|
chordal neural plate
,
blastopore
,
trigeminal placode
,
anterior neural fold
,
otic region
|
six1
|
|
laevis
|
NF stage 17
|
pre-chordal neural plate
,
neural plate
,
anterior neural fold
|
sox3
|
|
laevis
|
NF stage 17
|
trigeminal placode
,
neural plate
|
neurog2
|
|
laevis
|
NF stage 17
|
chordal neural plate
,
anterior neural fold
,
otic region
|
tubb2b
|
|
laevis
|
NF stage 17
|
chordal neural plate
,
neuron
,
otic region
|
neurod1
|
|
laevis
|
NF stage 17
|
chordal neural plate
,
otic region
|
cdknx
|
|
laevis
|
NF stage 17
|
chordal neural plate
,
anterior neural fold
|
|
|
Fig. 3. Effects of Eya1 and Six1 overexpression on markers of neurogenesis and placodal ectoderm at neural plate stages. Embryos after unilateral injection (lower half) of Eya1 (A–J), Six1 (K–N) or Eya1+Six1 (O) mRNA. In all cases, where myc-GFP was co-injected as lineage tracer (A–D, F–I, K, L, N, O), embryos are shown superimposed with green fluorescent channel. Black asterisks indicate reductions in the placodal or non-neural ectoderm. Arrows mark increased or ectopic marker gene expression in the placodal or non-neural ectoderm; black arrowhead identifies deformed blastopore due to gastrulation defects (O). For Sox3 (C, D) and p27Xic1 (I, J) after Eya1 injections two different phenotypes are depicted. |
|
Display additional annotations [+]
Gene |
Clone |
Species |
Stages |
Anatomy |
sox3
|
|
laevis
|
NF stage 16
|
midbrain
,
hindbrain
,
spinal cord
,
epibranchial placode
|
neurog2
|
|
laevis
|
NF stage 14
|
chordal neural plate
,
trigeminal placode
,
neuron
,
otic region
|
neurog2
|
|
laevis
|
NF stage 26
|
midbrain
,
hindbrain
,
spinal cord
,
trigeminal ganglion
,
lateral line placode
|
tubb2b
|
|
laevis
|
NF stage 14
|
chordal neural plate
,
trigeminal placode
,
neuron
|
tubb2b
|
|
laevis
|
NF stage 26
|
forebrain
,
midbrain
,
hindbrain
,
spinal cord
,
trigeminal ganglion
|
neurod1
|
|
laevis
|
NF stage 14
|
chordal neural plate
,
trigeminal placode
,
neuron
|
neurod1
|
|
laevis
|
NF stage 26
|
olfactory placode
,
otic vesicle
,
epibranchial placode
,
lateral line placode
|
neurod1
|
|
laevis
|
NF stage 16
|
trigeminal placode
|
cdknx
|
|
laevis
|
NF stage 14
|
chordal neural plate
|
|
|
Fig. 11. Role of SoxB1 genes in placodal neurogenesis. (A) Immunoblot showing that Sox2MO and Sox3MO specifically and efficiently block protein synthesis from co-injected utr-Sox2 and utr-Sox3 mRNA, respectively. (B–E, F–I) Effects of unilateral injection (lower half) of Sox2+Sox3MO (B–E), or Sox3 mRNA (F–I) on various neurogenic markers at neural plate stage. Embryos are shown superimposed with green fluorescent channel to reveal distribution of myc-GFP co-injected as lineage tracer. Asterisks indicate reductions, while arrows mark increased or ectopic marker gene expression in the placodal or non-neural ectoderm. (J–L) Effects of unilateral injection of GR–Sox3 and DEX activation at stage 16–18 on neurogenic markers at tail bud stages (J1–L1: injected side; J2–L2: control side). Asterisks indicate reductions in the placodal or non-neural ectoderm. (M, N) Ectopic neuronal differentiation (arrows) after injection of GR–Eya1+GR–Six1 mRNAs (M1: inj. side; M2: control side) and DEX activation at stage 16–18 is significantly reduced after co-injection of Sox2/Sox3 MO (M3: inj. side; M4: control side; N: quantitation, χ2 test; : p < 0.001). (O–R) Ectopic expression (arrows) of Sox3 (O1, O3: inj. side; O2, O4: control side) and Sox2 (P1, P3: inj. side; P2, P4: control side) but not of NeuroD (Q1, Q3: inj. side; Q2, Q4: control side) and p27Xic1 after injection of GR–Eya1+GR–Six1 mRNAs and DEX activation at stage 16–18 persists and is more extensive with CHX treatment (R: quantitation; χ2 test; : p < 0.05, : p < 0.001). |