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.