ZFIN ID: ZDB-FIG-050307-4
Amoyel et al., 2005 - Wnt1 regulates neurogenesis and mediates lateral inhibition of boundary cell specification in the zebrafish hindbrain. Development (Cambridge, England)   132(4):775-785 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Term:
Stage Range: 14-19 somites to Prim-5
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: 20-25 somites to Prim-5

Fig. 4 Regulation of neurogenesis in the hindbrain by Wnt signalling. In situ hybridisation or immunocytochemistry was carried out to detect expression of probe/antigen, as indicated at the top right of each panel. (A-D) Time-course of the effect of wnt1 knockdown on deltaD expression; no effect is observed at 18 hours, whereas expression is much reduced at 19.5 hours (8/10 embryos). (E,F) Neurogenesis is reduced in wnt1 MO-injected embryos at 24 hours, as seen by detecting p27xic1-a (p27) expression (20/20 embryos). (G-P) Knockdown of wnt1 (82/86 embryos), tcf3b (50/55 embryos) and rfng (28/37 embryos) all cause downregulation of delta and proneural gene expression at 24 hours. A greater decrease in deltaA expression occurs following knockdown of tcf3b than wnt1 (compare G and H). Notably, neurogenesis is less affected by these knockdowns in r4 (F-H,J-L,N,O), r7 and the spinal cord (arrows in N and O indicate the r6/r7 boundary). (P) Double asha/ashb knockdown decreases deltaD expression. Note that in knockdowns that disrupt neurogenesis, the hindbrain does not broaden and appears to be at an earlier morphological stage due to decreased cell proliferation and major depletion of the mantle zone of postmitotic neurons. (Q-X) Activating Wnt signalling causes ectopic neurogenesis. Hu expression in uninjected (Q), wnt MO injected (R), cWnt1 injected (S,T), stabilised ß-catenin injected (U,V) or double wnt MO and ß-catenin injected embryos (W,X). In T, V and X, expressing cells are green (co-injection of GFP), and r3 and r5 are blue (EphA4). Embryos overexpressing Wnt1 have more neurons than uninjected embryos (compare Q and S). ß-Catenin can cause ectopic neuronal differentiation in expressing cells in uninjected embryos (arrows in U point to ectopic Hu-positive cells that express ß-catenin in V), and in wnt1 MO embryos (compare left and right sides in W; more neurons are present on the left, where most ß-catenin expressing cells are present, X). Scale bars: in A, 100 µm for A-P; in Q, 50 µm for Q-X

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
ascl1b WT standard conditions Prim-5 hindbrain ISH
WT + MO1-rfng + MO2-rfng standard conditions Prim-5 hindbrain ISH
WT + MO1-tcf7l1b standard conditions Prim-5 hindbrain ISH
WT + MO1-wnt1 standard conditions Prim-5 hindbrain ISH
cdkn1ca WT standard conditions Prim-5 hindbrain ISH
WT + MO1-wnt1 standard conditions Prim-5 hindbrain ISH
dla WT + MO1-tcf7l1b standard conditions Prim-5 hindbrain ISH
WT + MO1-wnt1 standard conditions Prim-5 hindbrain ISH
dld WT standard conditions 14-19 somites hindbrain ISH
20-25 somites hindbrain ISH
WT + MO1-ascl1b + MO3-ascl1a standard conditions Prim-5 hindbrain ISH
WT + MO1-wnt1 standard conditions 14-19 somites hindbrain ISH
20-25 somites hindbrain ISH
epha4a WT standard conditions Prim-5 hindbrain IHC
WT + MO1-wnt1 standard conditions Prim-5 hindbrain ISH
neurog1 WT standard conditions Prim-5 hindbrain ISH
WT + MO1-tcf7l1b standard conditions Prim-5 hindbrain ISH
WT + MO1-wnt1 standard conditions Prim-5 hindbrain ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-ascl1b + MO3-ascl1a standard conditions Prim-5 cell proliferation in hindbrain disrupted, abnormal
Prim-5 hindbrain decreased width, abnormal
Prim-5 hindbrain morphogenesis delayed, abnormal
Prim-5 neurogenesis disrupted, abnormal
WT + MO1-rfng + MO2-rfng standard conditions Prim-5 cell proliferation in hindbrain disrupted, abnormal
Prim-5 hindbrain decreased width, abnormal
Prim-5 hindbrain morphogenesis delayed, abnormal
Prim-5 neurogenesis disrupted, abnormal
WT + MO1-tcf7l1b standard conditions Prim-5 cell proliferation in hindbrain disrupted, abnormal
Prim-5 hindbrain decreased width, abnormal
Prim-5 hindbrain morphogenesis delayed, abnormal
Prim-5 neurogenesis disrupted, abnormal
WT + MO1-wnt1 standard conditions 20-25 somites neurogenesis disrupted, abnormal
Prim-5 cell proliferation in hindbrain disrupted, abnormal
Prim-5 hindbrain decreased width, abnormal
Prim-5 hindbrain morphogenesis delayed, abnormal
Prim-5 neurogenesis disrupted, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development