ZFIN ID: ZDB-FIG-160526-16
Remédio et al., 2016 - Diverging roles for Lrp4 and Wnt signaling in neuromuscular synapse development during evolution. Genes & Development   30:1058-69 Full text @ Genes & Dev.
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Anatomical Terms:
Stage Range: 20-25 somites to Prim-25
PHENOTYPE:
Fish:
Observed In:
Stage Range: 20-25 somites to Prim-5

Fig. 6

Zebrafish Lrp4 is required for synapse formation but not AChR prepatterning. (A) Sequences of wild-type zebrafish lrp4 exon 9 and sequences of lrp4p184 and lrp4p185, two frameshift alleles that were generated using TALENs targeting exon 9. (B) The protein domain structure of wild-type zebrafish Lrp4 (1899 amino acids) and the predicted structure of lrp4p184 (331 amino acids), which resembles the mouse mitt allele and is predicted to lack the β-propeller domains, the EGF-like domains, the transmembrane domain, and the intracellular region. (C,C′) Lateral views of wild-type and lrp4 mutant zebrafish embryos at the prepatterning stage (19.5 h post-fertilization [hpf]) stained for AChRs (red) and motor axons (green). Lrp4 mutant embryos show no reduction or defect in AChR prepatterning. n = 50 out of 50 embryos. (D,D′) Lateral views of wild-type and lrp4 mutant zebrafish embryos at the synapse formation stage (26 hpf) stained for AChRs (red) and motor axons (green). In contrast to the prepatterning stage, lrp4 mutant embryos show a significant reduction in AChRs clustered beneath the motor axon terminals, with only a few “hot spot” AChR clusters remaining at the horizontal myoseptum (white arrowheads). n = 50 out of 50 embryos. (E,E′) Lateral views of in situ hybridizations performed on wild-type zebrafish embryos at the prepatterning stage (E) and synapse formation stage (E′). Lrp4 mRNA expression is undetectable at the prepatterning stage (E and zoomed panel) but robustly expressed at later stages (E′ and zoomed panel), consistent with a requirement for zebrafish Lrp4 during synapse formation but not during prepatterning.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
lrp4 p149Tg standard conditions 20-25 somites Not Detected adaxial cell ISH
Prim-25 muscle pioneer ISH
mCherry p149Tg standard conditions 20-25 somites adaxial cell IHC
Prim-25 muscle pioneer IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
znp-1 IHC WT standard conditions 20-25 somites motor neuron
IHC 20-25 somites motor neuron axon
IHC Prim-5 motor neuron
IHC Prim-5 motor neuron axon
IHC lrp4p184/p184 standard conditions 20-25 somites motor neuron
IHC 20-25 somites motor neuron axon
IHC Prim-5 motor neuron
IHC Prim-5 motor neuron axon
Phenotype Details
Fish Conditions Stage Phenotype
lrp4p184/p184 standard conditions 20-25 somites muscle acetylcholine-gated channel complex amount, normal
Prim-5 establishment of synaptic specificity at neuromuscular junction decreased occurrence, abnormal
Prim-5 muscle acetylcholine-gated channel complex decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Genes & Development for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Genes & Dev.