Figure 6.
CCAP neuron ablation causes PNS neuron dendrite pruning defects. (A–D′) Effect of CCAP neuron ablation on ecdysone target gene expression in c4da neurons. C4da neurons were labeled by ppk-CD4::tdtomato or ppk-CD4::tdGFP in control animals (CCAP > GFP, CCAP > lacZ) or in animals with ablated CCAP neurons (CCAP > hid), and immunofluorescence of Sox14 and Mical was performed at 2 h APF. (A–B′) Sox14 expression in CCAP > GFP controls (A and A′) and in CCAP-ablated animals (B and B′). (C–D′) Mical expression in CCAP > lacZ controls (C and C′) and in CCAP-ablated animals (D and D′). (E–H) Dendrite severing defects in lateroventral and ventral c4da neurons upon ablation of CCAP neurons. C4da neurons were labeled by ppk-CD4::tdtomato. Shown are hemisegments of pupae of the indicated genotypes at 18 h APF. Positions of the dorsal (d), lateroventral (v') and ventral (v) c4da neurons are indicated. (E) Control animal expressing GFP under the control of CCAP-GAL4. (F) Animal expressing UAS-hid under CCAP-GAL4 to ablate CCAP neurons. (G) Penetrance of dendrite severing defects in E and F as shown by the percentage of c4da neurons with unpruned, attached dendrites at 18 h APF. n.s., not significant, *P < 0.05, ***P < 0.0005, two-tailed Fisher’s exact test (N = 29–68). (H) Severity of dendrite severing defects in E and F as assessed by number of primary and secondary dendrites attached to the soma. Values are mean ± SEM, n.s., not significant, *P < 0.05, ***P < 0.0005, Wilcoxon’s test. (I–K) C1da neuron pruning defects upon CCAP neuron ablation. Multidendritic sensory neurons were labeled by expression of CD8::GFP under the control of nsyb-lexA. C1da neurons were identified by cell body and dendritic morphology. Cell body positions of the c1da neuron ddaD are marked by arrows. (I) Dorsal sensory neurons in control animals carrying CCAP-GAL4. (J) Dorsal sensory neurons in animal with ablated CCAP neurons. (K) Quantification of dendrite severing defects as shown by the number of primary and secondary dendrites attached to the ddaD cell body at 18 h APF. Values are mean ± SEM, ***P < 0.0005, Wilcoxon test (N = 39, 25). Scale bars are 10 μm in A and C, 100 μm in E, and 50 μm in I.

CCAP neuron ablation causes PNS neuron dendrite pruning defects. (A–D′) Effect of CCAP neuron ablation on ecdysone target gene expression in c4da neurons. C4da neurons were labeled by ppk-CD4::tdtomato or ppk-CD4::tdGFP in control animals (CCAP > GFP, CCAP > lacZ) or in animals with ablated CCAP neurons (CCAP > hid), and immunofluorescence of Sox14 and Mical was performed at 2 h APF. (A–B′) Sox14 expression in CCAP > GFP controls (A and A′) and in CCAP-ablated animals (B and B′). (C–D′) Mical expression in CCAP > lacZ controls (C and C′) and in CCAP-ablated animals (D and D′). (E–H) Dendrite severing defects in lateroventral and ventral c4da neurons upon ablation of CCAP neurons. C4da neurons were labeled by ppk-CD4::tdtomato. Shown are hemisegments of pupae of the indicated genotypes at 18 h APF. Positions of the dorsal (d), lateroventral (v') and ventral (v) c4da neurons are indicated. (E) Control animal expressing GFP under the control of CCAP-GAL4. (F) Animal expressing UAS-hid under CCAP-GAL4 to ablate CCAP neurons. (G) Penetrance of dendrite severing defects in E and F as shown by the percentage of c4da neurons with unpruned, attached dendrites at 18 h APF. n.s., not significant, *P < 0.05, ***P < 0.0005, two-tailed Fisher’s exact test (N = 29–68). (H) Severity of dendrite severing defects in E and F as assessed by number of primary and secondary dendrites attached to the soma. Values are mean ± SEM, n.s., not significant, *P < 0.05, ***P < 0.0005, Wilcoxon’s test. (I–K) C1da neuron pruning defects upon CCAP neuron ablation. Multidendritic sensory neurons were labeled by expression of CD8::GFP under the control of nsyb-lexA. C1da neurons were identified by cell body and dendritic morphology. Cell body positions of the c1da neuron ddaD are marked by arrows. (I) Dorsal sensory neurons in control animals carrying CCAP-GAL4. (J) Dorsal sensory neurons in animal with ablated CCAP neurons. (K) Quantification of dendrite severing defects as shown by the number of primary and secondary dendrites attached to the ddaD cell body at 18 h APF. Values are mean ± SEM, ***P < 0.0005, Wilcoxon test (N = 39, 25). Scale bars are 10 μm in A and C, 100 μm in E, and 50 μm in I.

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