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ZFIN ID: ZDB-FIG-170222-25
Fontenas et al., 2016 - Neuronal Ndrg4 Is Essential for Nodes of Ranvier Organization in Zebrafish. PLoS Genetics   12:e1006459 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Antibodies:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Protruding-mouth to Day 4
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Protruding-mouth to Day 4

Fig. 5

ndrg4 function is required for the expression of several genes that are essential for vesicle docking.

(A) qPCR showing a significant decrease in the expression of nsfa, syt1a, stxbp1b but not vamp2 in ndrg4 mutants with mRNA relative expression to ef1a. Controls from 3 different experiments have deliberately been set to 100 per cent. (n = 3 independent experiments of 30 embryos each). (B-D) Lateral views showing Snap25 expression in the PLLn axons at 4 dpf. Snap25 is visible all along the PLLn (arrows) in controls (B), but detected to a lesser extent in the ndrg4 mutants (C) and morphants (D) PLLn. (E-G) Lateral views of SV2 immunostaining, labeling synaptic vesicles at 4 dpf. (E) Synaptic vesicles are regularly distributed in the control PLLn (arrows) whereas they form agglomerates (arrowheads) in ndrg4 mutants (F) and morphants (G). Scale bars = 10μm. (H-K) Western blots showing a sharp decrease in Snap25 protein expression in ndrg4 mutants and morphants. Snap25 expression is significantly decreased in ndrg4 morphants (H,J) and mutants (I,K) with protein relative expression to β-actin. Controls from 3 different experiments have deliberately been set to 100 per cent. (n = 3 independent experiments of 30 embryos each). For the mutants study, the average of three groups of controls has been set to 100 per cent.

Gene Expression DetailsNo data available
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab-SV2 IHC WT control Day 4 posterior lateral line nerve synaptic vesicle
IHC WT + CRISPR1-ndrg4 standard conditions Day 4 posterior lateral line nerve synaptic vesicle
IHC WT + MO2-ndrg4 standard conditions Day 4 posterior lateral line nerve synaptic vesicle
Ab3-snap25 IHC WT control Day 4 posterior lateral line nerve axon
IHC WT + CRISPR1-ndrg4 standard conditions Day 4 posterior lateral line nerve axon
IHC WT + MO2-ndrg4 standard conditions Day 4 posterior lateral line nerve axon
WB WT control Protruding-mouth whole organism
WB WT + CRISPR1-ndrg4 standard conditions Protruding-mouth whole organism
WB WT + MO2-ndrg4 standard conditions Protruding-mouth whole organism
Phenotype Details
Fish Conditions Stage Phenotype
WT + CRISPR1-ndrg4 standard conditions Protruding-mouth whole organism Ab3-snap25 labeling decreased amount, abnormal
Day 4 posterior lateral line nerve axon Ab3-snap25 labeling decreased distribution, abnormal
Day 4 posterior lateral line nerve synaptic vesicle aggregated, abnormal
Day 4 posterior lateral line nerve synaptic vesicle ab-sv2 labeling spatial pattern, abnormal
WT + MO2-ndrg4 standard conditions Protruding-mouth whole organism Ab3-snap25 labeling decreased amount, abnormal
Day 4 posterior lateral line nerve axon Ab3-snap25 labeling decreased distribution, abnormal
Day 4 posterior lateral line nerve synaptic vesicle aggregated, abnormal
Day 4 posterior lateral line nerve synaptic vesicle ab-sv2 labeling spatial pattern, abnormal
Acknowledgments:
ZFIN wishes to thank the journal PLoS Genetics for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ PLoS Genet.