ZFIN ID: ZDB-FIG-110811-47
Webb et al., 2011 - The Enhancer of split transcription factor Her8a is a novel dimerisation partner for Her3 that controls anterior hindbrain neurogenesis in zebrafish. BMC Developmental Biology   11(1):27 Full text @ BMC Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage: 1-4 somites
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: 1-4 somites

Fig. 3

Like Her3, Her8a activity maintains the non-neurogenic areas of rhombomeres 2 and 4. A,B. neurog1 expression highlights the proneural clusters at the 3-somite stage (black arrows in A) and is eliminated upon her8a overexpression (B, embryo injected with her8a capped mRNA). Expression of tp63, which highlights the neural plate border (white arrowheads in A), is unchanged. C-F. Compared expression of neurog1 in control embryos (C) and embryos injected with her8aMO (D), her3MO (E) or both MOs (F) shows ectopic neurogenesis between the medial and lateral proneural clusters of r2 and r4 (blue arrows in D-F, compare with white arrowheads in C) when Her8a and/or Her3 activities are blocked. Few "ectopic" neurog1-positive can sometimes be found between the vcc and mnr2; this is however highly variable between individuals and observed in both control and morphant embryos. A-F are dorsal views of flat-mounted embryos, anterior left. Abbreviations: black arrows indicate proneural clusters: IN: presumptive interneurons, MN: presumptive motoneurons, r2: rhombomere 2, r4: rhombomere 4, r2l: lateral neurons of rhombomere 2, r4l: lateral neurons of rhombomere 4, SN: presumptive sensory neurons, vcc: ventro-caudal cluster. G. RT-PCR analysis of her8a expression (left and middle panels) in embryos injected with her8aMO1 ("mo1") and her8aMO2 ("mo2") versus control embryos ("ctr"). Low levels of full length, normally spliced her8a transcripts are detectable in morphants (left and middle panels, arrows) while abnormally spliced transcripts including all or part of intron 1 become produced (stars). Expression of βactin2, used as RT-PCR control, is indentical in all samples (right panel). The scheme at the top indicates the genomic structure of her8a, the position of exons (E, purple) and introns (black bars), the binding sites of her8aMO1 and MO2 (red), the position of RT-PCR primers (blue arrows) and the length of amplified wild-type products (excluding introns) (blue bars).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
neurog1 WT standard conditions 1-4 somites hindbrain neural keel interneuron ISH
1-4 somites hindbrain neural keel sensory neuron ISH
1-4 somites presumptive rhombomere 2 lateral region ISH
1-4 somites presumptive rhombomere 2 motor neuron ISH
1-4 somites presumptive rhombomere 4 lateral region ISH
1-4 somites presumptive rhombomere 4 motor neuron ISH
WT + MO1-her3 standard conditions 1-4 somites hindbrain neural keel interneuron ISH
1-4 somites hindbrain neural keel sensory neuron ISH
1-4 somites presumptive rhombomere 2 lateral region ISH
1-4 somites presumptive rhombomere 2 motor neuron ISH
1-4 somites presumptive rhombomere 4 lateral region ISH
1-4 somites presumptive rhombomere 4 motor neuron ISH
WT + MO1-her3 + MO3-her8a + MO4-her8a + MO5-her8a standard conditions 1-4 somites hindbrain neural keel interneuron ISH
1-4 somites hindbrain neural keel sensory neuron ISH
1-4 somites presumptive rhombomere 2 lateral region ISH
1-4 somites presumptive rhombomere 2 motor neuron ISH
1-4 somites presumptive rhombomere 4 lateral region ISH
1-4 somites presumptive rhombomere 4 motor neuron ISH
WT + MO3-her8a + MO4-her8a + MO5-her8a standard conditions 1-4 somites hindbrain neural keel interneuron ISH
1-4 somites hindbrain neural keel sensory neuron ISH
1-4 somites presumptive rhombomere 2 lateral region ISH
1-4 somites presumptive rhombomere 2 motor neuron ISH
1-4 somites presumptive rhombomere 4 lateral region ISH
1-4 somites presumptive rhombomere 4 motor neuron ISH
tp63 WT standard conditions 1-4 somites non neural ectoderm medial margin ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-her3 standard conditions 1-4 somites negative regulation of neurogenesis decreased occurrence, abnormal
1-4 somites rhombomere 2 neuron mislocalised, abnormal
1-4 somites rhombomere 4 neuron mislocalised, abnormal
WT + MO1-her3 + MO3-her8a + MO4-her8a + MO5-her8a standard conditions 1-4 somites negative regulation of neurogenesis decreased occurrence, abnormal
1-4 somites rhombomere 2 neuron mislocalised, abnormal
1-4 somites rhombomere 4 neuron mislocalised, abnormal
WT + MO3-her8a + MO4-her8a + MO5-her8a standard conditions 1-4 somites negative regulation of neurogenesis decreased occurrence, abnormal
1-4 somites rhombomere 2 neuron mislocalised, abnormal
1-4 somites rhombomere 4 neuron mislocalised, abnormal
Acknowledgments:
ZFIN wishes to thank the journal BMC Developmental Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ BMC Dev. Biol.