Figure 4.
PVD neuronal polarity and microtubule organization are not affected by postmitotic degradation of KNL-1. (A) Schematic showing the intracellular organization of the PVD anterior dendrite and axon. DMA-1 is a dendrite-specific receptor and RAB-3 is found in the membranes of synaptic vesicles (SV) that accumulate in the axon of PVD. Microtubules in the primary anterior dendrite are organized with minus ends facing the dendrite tip and the plus ends point toward the cell body. (B) Quantification of RAB-3 fluorescence intensity in the dendrite and axon of the PVD neuron in the indicated conditions. n represents the number of animals. Error bars represent mean ± 95% CI. P values from one-way ANOVA test followed by Tukey’s test. (C) Quantification of DMA-1 fluorescence intensity in the 4° dendrites and axons in indicated conditions. n represents number of animals. Error bars represent mean ± 95% CI. P values from Kruskal–Wallis test followed by Dunn’s test. (D) EB1EBP-2::GFP dynamics in the anterior primary dendrite. Image of the PVD primary anterior dendrite at the L4 larval stage (top). The green arrowheads indicate EB1EBP-2 comets in the dendrite. Scale bar, 5 µm. Below are kymographs of EB1EBP-2::GFP along the dendrite in control and KNL-1 DEG animals (bottom). Scale bar, 10 s and 10 μm, respectively. (E) Quantification of EB1EBP-2::GFP comet direction in control and KNL-1 DEG. n represents number of animals. Error bars represent mean ± 95% CI. P values from two-way ANOVA test followed by Šídák’s test. (F) Schematic of the split-GFP system used to express endogenous GFP::TBA-1 specifically in the PVD (top). The image below represents GFP::TBA-1 in the anterior dendrite and the cell body of the PVD. The red rectangle denotes the region of 15 μm along the primary dendrite where photobleaching was performed. Scale bar 5 µm. The graph depicts the fluorescence recovery of GFP::TBA-1 in control and KNL-1 DEG. n represents the number of animals. Lines and shaded regions represent mean ± 95% CI, respectively. ROI, region of interest. CB, cell body. (G–J) Quantification of EB1EBP-2::GFP comet dynamics in control and KNL-1 DEG. n represents the number of animals (G) and the number of comets (H–J) respectively. Error bars represent mean ± 95% CI. P values from unpaired t test (G) or Mann–Whitney test (H–J).

PVD neuronal polarity and microtubule organization are not affected by postmitotic degradation of KNL-1. (A) Schematic showing the intracellular organization of the PVD anterior dendrite and axon. DMA-1 is a dendrite-specific receptor and RAB-3 is found in the membranes of synaptic vesicles (SV) that accumulate in the axon of PVD. Microtubules in the primary anterior dendrite are organized with minus ends facing the dendrite tip and the plus ends point toward the cell body. (B) Quantification of RAB-3 fluorescence intensity in the dendrite and axon of the PVD neuron in the indicated conditions. n represents the number of animals. Error bars represent mean ± 95% CI. P values from one-way ANOVA test followed by Tukey’s test. (C) Quantification of DMA-1 fluorescence intensity in the 4° dendrites and axons in indicated conditions. n represents number of animals. Error bars represent mean ± 95% CI. P values from Kruskal–Wallis test followed by Dunn’s test. (D) EB1EBP-2::GFP dynamics in the anterior primary dendrite. Image of the PVD primary anterior dendrite at the L4 larval stage (top). The green arrowheads indicate EB1EBP-2 comets in the dendrite. Scale bar, 5 µm. Below are kymographs of EB1EBP-2::GFP along the dendrite in control and KNL-1 DEG animals (bottom). Scale bar, 10 s and 10 μm, respectively. (E) Quantification of EB1EBP-2::GFP comet direction in control and KNL-1 DEG. n represents number of animals. Error bars represent mean ± 95% CI. P values from two-way ANOVA test followed by Šídák’s test. (F) Schematic of the split-GFP system used to express endogenous GFP::TBA-1 specifically in the PVD (top). The image below represents GFP::TBA-1 in the anterior dendrite and the cell body of the PVD. The red rectangle denotes the region of 15 μm along the primary dendrite where photobleaching was performed. Scale bar 5 µm. The graph depicts the fluorescence recovery of GFP::TBA-1 in control and KNL-1 DEG. n represents the number of animals. Lines and shaded regions represent mean ± 95% CI, respectively. ROI, region of interest. CB, cell body. (G–J) Quantification of EB1EBP-2::GFP comet dynamics in control and KNL-1 DEG. n represents the number of animals (G) and the number of comets (H–J) respectively. Error bars represent mean ± 95% CI. P values from unpaired t test (G) or Mann–Whitney test (H–J).

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