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ZIRC
ZFIN ID: ZDB-FIG-160729-5
Lv et al., 2016 - Synaptic Ribbons Require Ribeye for Electron Density, Proper Synaptic Localization, and Recruitment of Calcium Channels. Cell Reports   15(12):2784-95 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Anatomical Terms:
Stage: Day 5
PHENOTYPE:
Fish:
Observed In:
Stage: Day 5

Fig. 2

Ribeye Is Dramatically Reduced from Neuromast Hair Cells of re[a(Δ10)/b(Δ7)] Double Homozygous Mutants 5 dpf

(A) Confocal images of Ribeye A (red) and post-synaptic density marker pan-MAGUK (green) staining 5 dpf in WT (left) and re[a(Δ10)/b(Δ7)] homozygous mutant (right) neuromast. Insets show magnified images centered around synapses. Note the close apposition of Ribeye a and pan-MAGUK in WT animals, whereas little Ribeye a staining is found near pan-MAGUK in the re[a(Δ10)/b(Δ7)] double-homozygous mutants.

(B) Ribeye b (red) and pan-MAGUK (green) antibody staining in 5-dpf WT (left) and re[a(Δ10)/b(Δ7)] homozygous mutant (right) neuromast.

(C) CtBP antibody staining (green) in 5-dpf WT (left) and re[a(Δ10)/b(Δ7)] homozygous mutant (right) neuromast. DAPI (blue) stains nucleus. Scale bars, 5 mm.

(D) The number of Ribeye a, MAGUK, or CtBP1 immunolabeled puncta per hair cell in 5- to 6-dpf WT and re[a(Δ10/b(Δ7) mutants. Each circle represents an individual neuromast (NM) within a larva. The number of puncta per hair cell was approximated by dividing the number of Ribeye puncta within an NM by the number of hair cells in the NM. Error bars indicate SEM. Unpaired t test; **p = 0.0075; *p = 0.0226; n.s., not significant.

(E) Integrated intensities per punctum of Ribeye a, MAGUK, and CtBP1. Bars represent the means; error bars indicate SEM. Mann-Whitney U test; ***p < 0.0001, for both mutant alleles.

(F) Cumulative histogram of distances between Ribeye a spots to their nearest pan-MAGUK spot in confocal micrographs for WT (red) and re[a(Δ10/b(Δ7)] double-homozygous mutants (black).

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
ctbp2a Ab1-ctbp2a WT standard conditions Day 5 neuromast hair cell ribbon synapse IHC
Ab1-ctbp2a ctbp2aya400/ya400; ctbp2lya401/ya401 standard conditions Day 5 neuromast hair cell ribbon synapse IHC
ctbp2l Ab2-ctbp2l WT standard conditions Day 5 neuromast hair cell ribbon synapse IHC
Ab2-ctbp2l ctbp2aya400/ya400; ctbp2lya401/ya401 standard conditions Day 5 Not Detected neuromast hair cell ribbon synapse IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-ctbp IHC WT standard conditions Day 5 neuromast hair cell ribbon synapse
IHC ctbp2aya400/ya400; ctbp2lya401/ya401 standard conditions Day 5 neuromast hair cell ribbon synapse
Ab1-pan-MAGUK IHC WT standard conditions Day 5 neuromast postsynaptic density
IHC ctbp2aya400/ya400; ctbp2lya401/ya401 standard conditions Day 5 neuromast postsynaptic density
Phenotype Details
Fish Conditions Stage Phenotype
ctbp2aya400/ya400; ctbp2lya401/ya401 standard conditions Day 5 neuromast hair cell ribbon synapse ctbp2l expression absent, abnormal
Day 5 neuromast hair cell ribbon synapse ab1-ctbp labeling decreased amount, abnormal
Day 5 neuromast hair cell ribbon synapse ctbp2a expression decreased amount, abnormal
Day 5 neuromast hair cell ribbon synapse ctbp2a expression mislocalised, abnormal
Day 5 neuromast postsynaptic density ab1-pan-maguk labeling decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Rep.