![]() Neuronal pruning is a commonly observed phenomenon for the developing nervous systems to ensure precise wiring of neural circuits. Two-way ANOVA with Tukey’s multiple comparison test was performed. In the group of small Spn-F puncta (diameter < 0.6 μm), there are significantly more Spn-F puncta colocalizing with Rab11-DN than with Rab11-CA or -wt. (E) Quantification of the number of colocalized Rab11/Spn-F puncta in each type of neuron. A colocalizing Spn-F punctum (B) and a non-colocalizing Spn-F punctum(D) are demonstrated in Rab11-DN-expressing neuron (A, arrow) and Rab11-CA-expressing neuron (C, arrow), respectively. Colocalizing puncta are defined by showing overlapping signal peaks in signal profile data (B), and non-colocalizing Spn-F puncta are defined by lacking overlapping signal peaks (D). (A-E) Spn-F-mCherry was co-expressed with Rab11-DN (dominant negative) (A), Rab11-CA (constitutively active) (C), in larval ddaC neurons for investigation of colocalization between Spn-F and Rab11 puncta. S4 Fig: The colocalization of Spn-F with dominant negative Rab11 in neurons. (G) Quantification of Rab11-GFP intensity in the soma of larval ddaC neurons with control (E) and Rab11-RNAi (F) using a two-tailed unpaired t test is shown (***, p<0.001.) for control, n = 9 for Rab11-RNAi, n = 10. (E’, F’) The neurons were visualized with ppk-GAL4 and UAS-mCD8RFP. (E, F) The signals of Rab11-GFP were decreased in the soma of ddaC neurons with Rab11-dsRNAs expression (F), compared to the wild-type cells with control Luciferase-dsRNAs expression (E). For wild type (wt), n = 20 for ppk-GAL4 and Rab11-dsRNAs, n = 43 for Rab11-dsRNAs, n = 20. (D) Quantification of the total length of unpruned dendrites in neurons at 16 h APF. The percentage of cells was determined by dividing the number of neurons with defective pruning by the total number of cells examined for each genotype for wild type (wt), n = 50 for ppk-GAL4 and Rab11-dsRNAs, n = 90 for Rab11-dsRNAs, n = 100. (C) Quantification of dendrite pruning phenotypes in neurons at 16 h APF. (A, B) At 16 h APF (after puparium formation), the dendrites were pruned in wild-type (wt) neurons (A), but remained attached to the neurons of Rab11 RNAi (RNA interference) mutants (B). S2 Fig: The knockdown of Rab11 expression specifically in C4da neurons results in dendrite pruning defects. ![]()
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