![]() While ddaC neurons overexpressing the UAS control construct pruned all the dendrites ( A), ddaC neurons overexpressing Patronin ( B) driven by Gal4 4-77 or GFP-Patronin ( C) driven by two copies of ppk-Gal4 at a higher level exhibited simple arbors at WP stage and consistent dendrite pruning defects at 16 hr APF. ( A–E) Live confocal images of ddaC neurons expressing mCD8-GFP driven by ppk-Gal4, Gal4 4-77 or ppk-CD4-tdGFP at WP and 16 hr APF. ***p<0.001 as assessed by two-tailed Student’s t-test. The number of samples (n) in each group is shown on the bars. Red arrowheads point to the ddaC somas, and blue arrowheads point to the ddaF somas. ( E) Like control ddaF neurons, patronin RNAi #1 ddaF neurons labelled by Gal4 109(2)80-driven mCD8-GFP were also eliminated by 16 hr APF. Red arrows point to the ddaD somas and open arrowheads to the ddaE somas. ( D) While the control class I ddaD/ddaE neurons labelled by Gal4 2-21-driven mCD8-GFP pruned their larval dendrites at 20 hr APF, patronin RNAi #1 ddaD/ddaE neurons failed to do so. Quantification of number of dendrite termini in control and patronin c9-c5 ddaC MARCM clones. ( C) Compared to FRT G13 control, patronin c9-c5 ddaC MARCM clones exhibited simple dendrite arbors at w元 stage. Quantification of the severing defects in control and mutant ddaC neurons at 32 hr APF. ( B) ddaC neurons overexpressing patronin RNAi #1 by ppk-Gal4 pruned away most of their dendrites at 32 hr APF. Quantification of the severing defects in control and mutant ddaC neurons at 16 hr APF. ( A) While the control neurons pruned all the dendrites at 16 hr APF, ddaC neurons overexpressing patronin RNAi #2 or patronin RNAi #3 by ppk-Gal4 exhibited dendrite pruning defects at 16 hr APF. ( A–E) Live confocal images of da neurons expressing mCD8-GFP at w3L, WP, 16 hr and 20 hr, 32 hr APF. melanogaster cell biology dendrite dendrite morphology developmental biology microtubule orientation minus end neuron pruning. The Reviewing Editor's assessment is that minor issues remain unresolved (see decision letter).ĭ. This article has been through an editorial process in which the authors decide how to respond to the issues raised during peer review. Thus, our study demonstrates that Patronin orients minus-end-out MT arrays in dendrites to promote dendrite-specific pruning mainly through antagonizing Klp10A activity. Consistently, attenuation of Klp10A MT depolymerase in patronin mutant neurons significantly restored minus-end-out MTs in dendrites and thereby rescued dendrite-pruning defects. Moreover, we show that both patronin knockdown and overexpression resulted in a drastic decrease of MT minus ends and a concomitant increase of plus-end-out MTs in ddaC dendrites, suggesting that Patronin stabilizes dendritic minus-end-out MTs. The CKK domain is important for Patronin's function in dendrite pruning. Here, we identified Patronin, a minus-end-binding protein, for its crucial and dose-sensitive role in ddaC dendrite pruning. However, a requirement of MT minus-end-binding proteins in dendrite-specific pruning remains completely unknown. ddaCs distribute the minus ends of microtubules (MTs) to dendrites but the plus ends to axons. Class IV ddaC neurons specifically prune larval dendrites without affecting axons during Drosophila metamorphosis.
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