ZFIN ID: ZDB-FIG-090320-73
Jing et al., 2009 - Wnt signals organize synaptic prepattern and axon guidance through the zebrafish unplugged/MuSK receptor. Neuron   61(5):721-733 Full text @ Neuron
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 14-19 somites to Prim-5
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: 14-19 somites to Prim-5

Fig. 3 wnt11r Is Critical for Axonal Guidance and AChR Prepatterning

(A) The splice morpholino (SP-MO) targets the splice donor site of the wnt11r exon 3 (red arrow), and MO-induced aberrant splicing is shown in red. RT-PCR analyses of uninjected and wnt11r SP-MO injected embryos (arrows indicate the position of PCR primers).

(B) Quantification of wnt11r MOs injected embryos. TL-MO, translation initiation mopholino. Per embryo, 20 hemisegments were analyzed; n = hemisegments. Results are expressed as the mean of multiple injection experiments ± SEM, (*p < 0.001, t test).

(C–L) Wild-type, unplugged, and wnt11r MO injected embryos at 27 hpf (C–H) and at the 20 somite stage (I–L), stained for motor axons (znp-1, green) and AChR clusters (α-BTX, red). (E and F) In contrast to wild-type, unplugged embryos display characteristic stalling (arrowhead) and branches (arrows) at the choice point and lack all AChR clusters. (G and H) Injection of wnt11r MO causes unplugged-like axonal stalling (arrowhead), branching (arrow), and a strong reduction of AChR prepatterning (K and L). Note that the size and intensity of neural AChR clusters is reduced in wnt11r 27 hpf morphants (H).

(M–Q) Time-lapse images of Hb9-GFP-labeled wild-type (M and O) and wnt11r morphant CaP and VaP axons (N, P, and Q) as they exit from the spinal cord (M and N) and as they reach the somitic choice point (O–Q). Asterisks indicate the cell body of interneurons. (M and O) Wild-type CaP and VaP neurons extended one growth cone (arrow). Note the broad area (brackets) the two defasciculated wnt11r morphants CaP/VaP growth cones occupy (arrowheads in [N], [P], and [Q]), compared to wild-type (M and O).

Scale bars, 50 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
chrna1 WT standard conditions 20-25 somites myotome acetylcholine-gated channel complex OTHR
Prim-5 myotome acetylcholine-gated channel complex OTHR
WT + MO3-wnt11 standard conditions 20-25 somites myotome acetylcholine-gated channel complex OTHR
Prim-5 myotome acetylcholine-gated channel complex OTHR
musktbr307/tbr307 standard conditions Prim-5 Not Detected myotome OTHR
GFP ml2Tg standard conditions 14-19 somites CaP motoneuron axon IFL
14-19 somites CaP motoneuron neuronal cell body IFL
14-19 somites VaP motor neuron axon IFL
14-19 somites VaP motor neuron neuronal cell body IFL
ml2Tg + MO3-wnt11 standard conditions 14-19 somites CaP motoneuron axon IFL
14-19 somites CaP motoneuron neuronal cell body IFL
14-19 somites VaP motor neuron axon IFL
14-19 somites VaP motor neuron neuronal cell body IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
znp-1 IHC WT standard conditions 20-25 somites motor neuron axon
IHC Prim-5 motor neuron axon
IHC WT + MO3-wnt11 standard conditions 20-25 somites motor neuron axon
IHC Prim-5 motor neuron axon
IHC musktbr307/tbr307 standard conditions Prim-5 motor neuron axon
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO3-wnt11 standard conditions 20-25 somites motor neuron axon guidance process quality, abnormal
20-25 somites skeletal muscle acetylcholine-gated channel clustering decreased occurrence, abnormal
Prim-5 motor neuron axon guidance process quality, abnormal
Prim-5 skeletal muscle acetylcholine-gated channel clustering decreased occurrence, abnormal
ml2Tg + MO3-wnt11 standard conditions 14-19 somites CaP motoneuron axon mislocalised, abnormal
14-19 somites motor neuron axon guidance process quality, abnormal
14-19 somites VaP motor neuron axon mislocalised, abnormal
musktbr307/tbr307 standard conditions Prim-5 motor neuron axon guidance process quality, abnormal
Prim-5 skeletal muscle acetylcholine-gated channel clustering arrested, abnormal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Neuron