The function of cytoplasmic dynein complex is required for Spn-F redistribution and dendrite pruning.

<p>(A-D) Immunofluorescence of Spn-F-GFP, Ik2-HA and nuclear DAPI staining in S2 cells (A, B), in S2 cells treated with colchicine (C), and in S2 cells treated with ciliobrevin D (D). (E'-J') The ddaC neurons were visualized with <i>ppk-GAL4</i> and <i>UAS-mCD8RFP</i>. The Spn-F-GFP displayed as puncta in the soma of ddaC neurons of wild type (E), <i>ctp</i>^<i>G0153</i> (F), and <i>Dhc-RNAi</i> (G) larvae. At 5 h APF, the Spn-F-GFP puncta were dispersed in ddaC neurons of wild-type pupae (H), but remained as puncta in both <i>ctp</i>^<i>G0153</i> (I) and <i>Dhc-RNAi</i> (J) pupae. The thicken axons (arrows in G’ and J’) were caused by reduction of functional dynein complexes in <i>Dhc-RNAi</i> neurons. (K-P) The ddaC neurons were marked with <i>ppk-GAL4</i> and <i>UAS-mCD8GFP</i>. At 16 h APF, the dendrites of ddaC neurons were pruned in wild-type (K), but not in <i>Dhc-RNAi</i> (L), or <i>Dhc</i>^<i>6-6</i><i>/Df</i> mutant (M) pupae. Df: deficiency line Df(3L)BSC436, which deletes <i>Dhc</i> gene. (N) The dendrites of ddaC cells were severed in the proximal dendrites (arrows), but not cleaned up in <i>ctp</i>^<i>G0153</i> mutant pupae at 16 h APF. At 12 h APF, the dendrites of ddaC neurons are all severed (arrows) in wild-type pupae (O), but remained attached to the soma of ddaC cells in <i>ctp</i>^<i>G0153</i> pupae (P). (Q) Quantitative analysis of dendrite pruning phenotypes in ddaC cells of <i>Dhc</i> mutants at 16 h APF. The percentage of ddaC neurons shows dendrite pruning defects among the total number of neurons examined. For wild-type control, n = 100; for <i>Dhc</i>^<i>6-6</i><i>/Dhc</i>^<i>6-10</i> mutants, n = 89; for <i>Dhc</i>^<i>6-6</i><i>/Df</i> mutants, n = 90; for <i>Dhc</i>^<i>6-10</i><i>/Df</i> mutants, n = 100. (R) Quantitative analysis of dendrite pruning phenotypes in ddaC cells of <i>Dhc</i> mutants with SpnF-8D expression at 16 h APF. The percentage of ddaC neurons shows dendrite pruning defects among the total number of neurons examined. For wild-type control, n = 100; for <i>Dhc</i>^<i>6-6</i><i>/Df</i> mutants, n = 130; for <i>Dhc</i>^<i>6-6</i><i>/Df</i> mutants rescued with <i>spnF-8D</i>, n = 110. Statistical analysis was performed with Chi-square test. N.S., not significant. Scale bars, 10 μm (A-J), 20 μm (K-P).