Subcellular distribution of Hrp48 in MB neurons.

<p>(A) Cell bodies of UAS-mCD8-GFP/+;;OK107-Gal4/+ adult (A) brains stained with anti-Hrp48 antibodies (white in A; red in A’) and GFP (green in A’). (B) Cell bodies of UAS-mCD8-GFP/+;UAS-Flag-Hrp48/+;OK107-Gal4/+ adult brain stained with anti-Flag antibodies (white in B; red in B’) and GFP (green in B’). Scale bar: 20μm. (C) MB lobes of UAS-mCD8-GFP/+;;OK107-Gal4/+ adult brains stained with anti-Hrp48 antibodies (white in C; red in C’) and GFP (green in C’). (D) MB lobe of UAS-mCD8-GFP/+;UAS-Flag-Hrp48/+;OK107-Gal4/+ adult brain stained with anti-Flag antibodies (white in D; red in D’) and GFP (green in D’). Scale bar: 20μm.</p

<i>hrp48</i> downregulation induces ectopic projection of MB dorsal lobes.

<p>(A,B). MB lobes of control (A) and <i>hrp48</i>^{<i>k10413</i>}/<i>hrp48</i>^<i>02647</i> (B) adult brains stained with anti-FasII antibodies. (C-E) MB lobes of control (C) or <i>hrp48</i>-RNAi (D,E) adult brains expressing mCD8-GFP (green), and stained with anti-FasII antibodies (red). Arrows in B,D,E point to overextended dorsal axonal branches. Precise genotypes: UAS-mCD8-GFP/+;;OK107-Gal4/+ (C); UAS-mCD8-GFP/UAS-RNAi-<i>hrp48</i>^{<i>#101555</i>};;OK107-Gal4/+ (D,E). Scale bar in A-E: 20μm. (F) Percentages of MBs exhibiting normal projections (<i>ie</i> dorsal branches stopping at the end of the dorsal lobe), or ectopic projections (as judged based on the αβ axon bundle).</p

<i>hrp48</i> downregulation results in MB lobe projection defects.

<p>(A-C) Mushroom Body (MB) lobes of control (A), <i>hrp48</i>^{<i>k10413</i>}/<i>hrp48</i>^{<i>k16203</i>} (B), or <i>hrp48</i>^{<i>k10413</i>}/<i>hrp48</i>^<i>02647</i> (C) adult brains stained with anti-FasciclinII (FasII) antibodies. FasII is strongly expressed in αβ neurons, weakly in γ neurons. (D-F) MB lobes of control (D) or <i>hrp48</i>-RNAi (E,F) adult brains expressing mCD8-GFP (green), and stained with anti-FasII antibodies (red). Representative examples of the asymmetric and truncated categories used for quantification are shown. Precise genotypes: UAS-mCD8-GFP/+;;OK107-Gal4/+ (D); UAS-mCD8-GFP/UAS-RNAi-<i>hrp48</i>^{<i>#101555</i>};;OK107-Gal4/+ (E,F); UAS-mCD8-GFP/+;UAS-RNAi-<i>hrp48</i>^{<i>#16041</i>}<i>/+</i>;OK107-Gal4/+ (G). Scale bar in A-F: 20μm. (G) Percentages of MBs exhibiting symmetric projections, asymmetric projections or truncated lobes (either α or β) in controls (<i>w</i>), <i>hrp48</i> transheterozygous combinations, and two independent RNAi lines (#16041 and #101555). Two controls were used for the rescue assay: UAS-mCD8-GFP/UAS-RNAi-<i>hrp48</i>^{<i>#101555</i>};UAS-GFP/+;OK107-Gal4/+ flies as a control of Gal4 titration, and UAS-mCD8-GFP/+;UAS-Flag-Hrp48/+;OK107-Gal4/+ flies to ensure that Hrp48 overexpression did not induce axon projection defects. Numbers represent numbers of scored hemispheres. Statistical comparison to the UAS-GFP control: ***, p<0.001 (χ² test).</p

<i>hrp48</i> locus and mutants.

<p>(A) Genomic organization of <i>hrp48</i> locus, and intron-exon structure of <i>hrp48</i> transcripts. Untranslated and coding regions are represented respectively as white and black boxes. Positions of the P-elements inserted into the l(2)02647, l(2)k16203, l(2)k10413 and l(2)k02814 strains are shown in blue. Sequences targeted by the RNAi lines #101555 and 16041 are represented in red. Position of the EMS point mutation found in the 7E17-8 line is indicated in green. This mutation generates a premature stop codon at position 312 (<a href="http://flybase.org/reports/FBrf0198712.html" target="_blank">http://flybase.org/reports/FBrf0198712.html</a>). Note that the transcript nomenclature follows that of Flybase (FB2015_01 release). For the sake of simplicity, alternative 3’UTRs are not represented in the scheme (see open dotted boxes). (B) Western-Blot of control (w) and mutant (<i>hrp48</i>^{<i>k10413</i>}/<i>hrp48</i>^<i>02647</i> and <i>hrp48</i>^{<i>k10413</i>}/<i>hrp48</i>^{<i>k16203</i>}) adult brain extracts probed with anti-Hrp48 (upper lane) and anti-Tubulin (Tub; lower lane) antibodies. Values shown at the bottom correspond to normalized Hrp48/Tubulin signal intensity ratios. (C) UAS-mCD8-GFP/+;;OK107-Gal4/+ (upper panel) and UAS-mCD8-GFP/UAS<i>-RNAi</i>-<i>hrp48</i>^#101555;;OK107-Gal4/+ (lower panel) larval brains stained with anti-Hrp48 antibodies (left, red in the overlay) and GFP (green in the overlay). Note the reduced Hrp48 levels observed in MB cell bodies upon RNAi expression. Scale bar: 10 μm.</p

<i>hrp48</i> is required to prevent overextension of MB dorsal axonal branches.

<p>(A, B) Axonal projections of wild-type (A) or <i>hrp48</i>^{<i>7E7-18</i>} (B) neuroblast (Nb) clones labelled by mCD8-GFP and generated using the MARCM technique. Arrow in B points to ectopically projecting axonal branches. Scale bar: 20μm. (C) Percentages of Nb clones exhibiting « normal » projections (<i>ie</i> dorsal branches stopping at the end of the dorsal lobe), or ectopic projections. Numbers represent numbers of scored Nb clones. Re-expression of a wild-type copy of Hrp48 (Flag-Hrp48) rescues the MARCM mutant phenotypes (***, p<0.001 in a χ² test). Precise genotypes: hsp-flp, UAS-mCD8-GFP/+; FRT40A/FRT40A tub-Gal80;;OK107-Gal4/+ (A); hsp-flp, UAS-mCD8-GFP/+; FRT40A, <i>hrp48</i>^{<i>7E7-18</i>}/ FRT40A tub-Gal80;;OK107-Gal4/+ (B).</p

Tabu search using variable amplitude for dimensioning an assembly/ disassembly production system

<div><p>The confined and crowded environment of developing brains imposes spatial constraints on neuronal cells that have evolved individual and collective strategies to optimize their growth. These include organizing neurons into populations extending their axons to common target territories. How individual axons interact with each other within such populations to optimize innervation is currently unclear and difficult to analyze experimentally <i>in vivo</i>. Here, we developed a stochastic model of 3D axon growth that takes into account spatial environmental constraints, physical interactions between neighboring axons, and branch formation. This general, predictive and robust model, when fed with parameters estimated on real neurons from the <i>Drosophila</i> brain, enabled the study of the mechanistic principles underlying the growth of axonal populations. First, it provided a novel explanation for the diversity of growth and branching patterns observed <i>in vivo</i> within populations of genetically identical neurons. Second, it uncovered that axon branching could be a strategy optimizing the overall growth of axons competing with others in contexts of high axonal density. The flexibility of this framework will make it possible to investigate the rules underlying axon growth and regeneration in the context of various neuronal populations.</p></div

Additional file 3 of Analysis of asymptomatic Drosophila models for ALS and SMA reveals convergent impact on functional protein complexes linked to neuro-muscular degeneration

Additional file 3. Supplementary Dataset file 3

Additional file 5 of Analysis of asymptomatic Drosophila models for ALS and SMA reveals convergent impact on functional protein complexes linked to neuro-muscular degeneration

Additional file 5. Supplementary Dataset file 5

Additional file 1 of Analysis of asymptomatic Drosophila models for ALS and SMA reveals convergent impact on functional protein complexes linked to neuro-muscular degeneration

Additional file 1. Supplementary Dataset file 1

Additional file 2 of Analysis of asymptomatic Drosophila models for ALS and SMA reveals convergent impact on functional protein complexes linked to neuro-muscular degeneration

Additional file 2. Supplementary Dataset file 2