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. 2019 Jun 4;116(25):12327–12336. doi: 10.1073/pnas.1820928116

Fig. 3.

Fig. 3.

Testing heterochronic mutants for DA9 neuron GFP::RAB-3 polarity defects at early developmental stages. (A) A diagram depicting the timeline of the DA9 neuron development. Phenotypes of heterochronic mutants were scored at larval stages L2, L3, and L4. (B and C) Localization of GFP::RAB-3 in wild-type and lin-29(n482) animals at the L2 larval stage. In both wild-type and lin-29(n482) mutant animals, RAB-3 is localized at the dorsal side in the axonic region of DA9. (B) Schematic diagrams showing phenotypes are displayed above each image. (Scale bars, 10 μm.) (C) Quantification of the localization of GFP::RAB-3 of wild-type and lin-29(n482) animals at L2 larval stage. Images were generated by a confocal microscope. n = 37–47 worms per genotype. n.s is not significant. Error bars are SEM. (D and E) Animals expressing GFP::RAB-3 specifically in the DA9 (wyIs85) in wild type, dre-1(dh99), and lin-29(n482) mutants at L3 larval stage. In dre-1(dh99) mutants there is localization of GFP::RAB-3 in the normally asynaptic domain of DA9 as shown in D. Arrows indicate ectopic GFP::RAB-3 in DA9. Linescans were generated by a confocal microscope. Schematic diagrams showing phenotypes are displayed above each image. (Scale bars, 10 μm.) (E) Quantification of the GFP::RAB-3 localization at the asynaptic domain at L3 larval stage. n = 26–38 worms per genotype. *P < 0.05, two-tailed Student’s t test. Error bars are SEM. (F–H) Localization of RAB-3 puncta in wild type, dre-1(dh99), and lin-29(n482) mutants at L4 larval stage. (F) Representative confocal images of animals expressing the synaptic vesicle marker GFP::RAB-3 in wild type, dre-1(dh99), and lin-29(n482) mutants. Arrows indicate the ectopic GFP::RAB-3 puncta at the asynaptic domain of DA9. (Scale bars, 10 μm.) Schematic diagrams showing phenotypes are displayed above each image. (G) Quantification of GFP::RAB-3 localization along the DA9 in wild type and dre-1(dh99) at L4 larval stage. n = 54–60 worms per genotype. *P < 0.05, two-tailed Student’s t test. Error bars are SEM. (H) Quantification of GFP::RAB-3 localization along the DA9 in wild type and lin-29(n482) at L4 larval stage. n = 60 worms per genotype. n.s is not significant, two-tailed Student’s t test. Error bars are SEM.