ZFIN ID: ZDB-FIG-150609-7
Love et al., 2015 - Rest represses maturation within migrating facial branchiomotor neurons. Developmental Biology   401(2):220-35 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibody:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Long-pec to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Long-pec to Protruding-mouth

Fig. 1

Maternal rest is required for proper FBMN migration. (A) PCR genotyping of mating crosses to produce rest zygotic and maternal–zygotic (MZ) mutant embryos. The restsbu29 allele introduces a 7 bp deletion resulting in a smaller PCR product (red arrowhead). Homozygous mutant females (red box) produced eggs lacking maternal rest transcript. (B–F) Average intensity projection confocal images (dorsal views) of Tg(islet1:GFP); restsbu29 mutant embryos at 48 hpf, immunolabeled with EphA4a to allow visualization of r5 (red). Scale bar=20 µm. Decreasing levels of rest from B to F correlate with increasing disruption in FBMN migration. (B′–F′) Pie charts represent the average number of FBMNs counted in r4 (black), r5 (checkered), and r6 (white) as a percentage of total FBMNs in rest mutant embryos. The total number of FBMNs was not significantly different in embryos of different genotypes. (G) Statistical analysis of FBMN migration in restsbu29 mutants. The percentages of FBMNs that fully migrate to r6 (white) were compared using unpaired t test. Parentheses indicate number of embryos analyzed per group. Mean±SEM; *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001; n.s.=not significant. (H and I) Tg(islet1:GFP) embryos co-injected with both splice- and translation-blocking morpholinos targeted against Rest recapitulate the MZrestsbu29/sbu29 mutant phenotype at 48 hpf. Scale bar=20 µm. (J and K) MZrestsbu29/sbu29 FBMNs fail to reach r6 at 72 hpf, indicating that the mutant phenotype is not merely a consequence of developmental delay.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
GFP restsbu29/+; rw0Tg control Long-pec facial nerve motor nucleus motor neuron IHC
restsbu29/+; rw0Tg control Long-pec facial nerve motor nucleus motor neuron IHC
restsbu29/sbu29; rw0Tg control Long-pec facial nerve motor nucleus motor neuron IHC
restsbu29/sbu29; rw0Tg control Long-pec facial nerve motor nucleus motor neuron IHC
restsbu29/sbu29; rw0Tg control Protruding-mouth facial nerve motor nucleus motor neuron IHC
rw0Tg control Long-pec facial nerve motor nucleus motor neuron IHC
Protruding-mouth facial nerve motor nucleus motor neuron IHC
rw0Tg + MO1-rest + MO5-rest control Long-pec facial nerve motor nucleus motor neuron IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
ab1-epha4a IHC restsbu29/+; rw0Tg control Long-pec rhombomere 5
IHC restsbu29/+; rw0Tg control Long-pec rhombomere 5
IHC restsbu29/sbu29; rw0Tg control Long-pec rhombomere 5
IHC restsbu29/sbu29; rw0Tg control Long-pec rhombomere 5
IHC restsbu29/sbu29; rw0Tg control Protruding-mouth rhombomere 5
IHC rw0Tg control Long-pec rhombomere 5
IHC Protruding-mouth rhombomere 5
IHC rw0Tg + MO1-rest + MO5-rest control Long-pec rhombomere 5
Phenotype Details
Fish Conditions Stage Phenotype
restsbu29/+; rw0Tg control Long-pec facial nerve motor nucleus motor neuron displaced to rhombomere 4, abnormal
restsbu29/+; rw0Tg control Long-pec facial nerve motor nucleus motor neuron displaced to rhombomere 4, abnormal
restsbu29/+; rw0Tg control Long-pec facial nerve motor nucleus motor neuron displaced to rhombomere 5, abnormal
restsbu29/+; rw0Tg control Long-pec facial nerve motor nucleus motor neuron displaced to rhombomere 5, abnormal
restsbu29/+; rw0Tg control Long-pec facial nerve motor nucleus motor neuron migration decreased occurrence, abnormal
restsbu29/+; rw0Tg control Long-pec facial nerve motor nucleus motor neuron migration decreased occurrence, abnormal
restsbu29/+; rw0Tg control Long-pec rhombomere 6 has fewer parts of type facial nerve motor nucleus motor neuron, abnormal
restsbu29/+; rw0Tg control Long-pec rhombomere 6 has fewer parts of type facial nerve motor nucleus motor neuron, abnormal
restsbu29/sbu29; rw0Tg control Long-pec facial nerve motor nucleus motor neuron displaced to rhombomere 4, abnormal
restsbu29/sbu29; rw0Tg control Long-pec facial nerve motor nucleus motor neuron displaced to rhombomere 4, abnormal
restsbu29/sbu29; rw0Tg control Long-pec facial nerve motor nucleus motor neuron displaced to rhombomere 5, abnormal
restsbu29/sbu29; rw0Tg control Long-pec facial nerve motor nucleus motor neuron displaced to rhombomere 5, abnormal
restsbu29/sbu29; rw0Tg control Long-pec facial nerve motor nucleus motor neuron migration decreased occurrence, abnormal
restsbu29/sbu29; rw0Tg control Long-pec facial nerve motor nucleus motor neuron migration decreased occurrence, abnormal
restsbu29/sbu29; rw0Tg control Long-pec rhombomere 6 has fewer parts of type facial nerve motor nucleus motor neuron, abnormal
restsbu29/sbu29; rw0Tg control Long-pec rhombomere 6 has fewer parts of type facial nerve motor nucleus motor neuron, abnormal
restsbu29/sbu29; rw0Tg control Protruding-mouth facial nerve motor nucleus motor neuron displaced to rhombomere 4, abnormal
Protruding-mouth facial nerve motor nucleus motor neuron displaced to rhombomere 5, abnormal
Protruding-mouth facial nerve motor nucleus motor neuron migration decreased occurrence, abnormal
Protruding-mouth rhombomere 6 has fewer parts of type facial nerve motor nucleus motor neuron, abnormal
rw0Tg + MO1-rest + MO5-rest control Long-pec facial nerve motor nucleus motor neuron displaced to rhombomere 4, abnormal
Long-pec facial nerve motor nucleus motor neuron displaced to rhombomere 5, abnormal
Long-pec facial nerve motor nucleus motor neuron migration decreased occurrence, abnormal
Long-pec rhombomere 6 has fewer parts of type facial nerve motor nucleus motor neuron, abnormal
Acknowledgments:
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Reprinted from Developmental Biology, 401(2), Love, C.E., Prince, V.E., Rest represses maturation within migrating facial branchiomotor neurons, 220-35, Copyright (2015) with permission from Elsevier. Full text @ Dev. Biol.