A neural progenitor mitotic wave is required for asynchronous axon outgrowth and morphology
- Sorbonne Université, CNRS, Laboratoire de Biologie du Développement - Institut de Biologie Paris Seine (LBD-IBPS), Cell cycle and cell determination Team, France
- Sorbonne Université, INSERM, NutriOmics Research Unit, France
Figures
Figure 1 with 3 supplements
G2-arrested SOP cells of the neuroepithelium resume division in a temporal wave.
(A) Four frames of a time-lapse recording of a control Drosophila melanogaster pupa from Video 1. The first three rows of cells in the most dorsal part of the thorax are indicated as R1, R2, and R3 …
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Figure 1—source data 1
Related to Figure 1 Raw data.
- https://cdn.elifesciences.org/articles/75746/elife-75746-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
Illustrations of individual mitotic waves in control and scaBP2 mutant.
Heatmaps of nine nota showing the time of SOP division in control (A) and scaBP2 pupae (B). Circles represent SOP cells arranged in rows, anterior to the left. The relative time of division is …
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Figure 1—figure supplement 1—source data 1
Related to Figure 1—figure supplement 1.
- https://cdn.elifesciences.org/articles/75746/elife-75746-fig1-figsupp1-data1-v2.xlsx
Figure 1—figure supplement 2
The mitotic wave origin spreads out in several genetic contexts.
Location of the first dividing SOP (SOP0) in rows 1–3 in scaBP2 flies (B, n = 9) and its control, neur> GFP flies, (A, n = 9), after overexpression of rac1N17 (E, n = 6) or RNAi-sca (F, n = 9) and …
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Figure 1—figure supplement 2—source data 1
Related to Figure 1—figure supplement 2 Raw data.
- https://cdn.elifesciences.org/articles/75746/elife-75746-fig1-figsupp2-data1-v2.xlsx
Figure 1—figure supplement 3
Reduced mitotic waves were better fitted with low values of the inhibitory parameter μ.
(A–E) The time of SOP cell division (mean time ± SEM) in row 1 (R1), row 2 (R2), and row 3 (R3) is plotted relative to the position and time of division of the first cell to divide in each row. The …
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Figure 1—figure supplement 3—source data 1
Related to Figure 1—figure supplement 3 Raw data.
- https://cdn.elifesciences.org/articles/75746/elife-75746-fig1-figsupp3-data1-v2.xlsx
Figure 2 with 5 supplements
The SOP mitotic wave is affected in conditions where cell protrusions are reduced.
(A) Frames from live recordings showing cell protrusions of SOPs in control (left) and rac1N17 (right) pupae. tubGal80ts was used to conditionally express Gal4 and thus rac1N17 by maintaining flies …
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Figure 2—source data 1
Related to Figure 2 Raw data.
- https://cdn.elifesciences.org/articles/75746/elife-75746-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
Illustrations of individual mitotic waves after reduction of cell protrusions by overexpression of Rac1N17 and after conditional inactivation of sca.
Heatmaps of six nota showing the time of SOP division in control neur> ph (PLCδ)::RFP/+; pnr> Gal4 tub> Gal80ts/+ pupae (A), when cell protrusions were reduced in neur> ph (PLCδ)::RFP/+; pnr> Gal4 …
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Figure 2—figure supplement 1—source data 1
Related to Figure 2—figure supplement 1 Raw data.
- https://cdn.elifesciences.org/articles/75746/elife-75746-fig2-figsupp1-data1-v2.xlsx
Figure 2—figure supplement 2
Sca is transiently expressed in the notum.
Images of nota in sca::GFSTF protein-trap flies at different developmental times. Sca::GFSTF protein was transiently detected as multiple foci in sensory organ cells. Scale bar, 20 µm.
Figure 2—video 1
Time lapse recording showing the dynamic of protrusions in SOP cells in a control pupa.
The membrane was visualised using a membrane tethered red fluorescent protein (ph-RFP). Each frame was obtained by combining a z-stack (composed of 15 optical sections separated by 1 µm) acquired …
Figure 2—video 2
Time lapse recording showing the dynamics of protrusions of SOPs in pupa overexpressing the negative form of Rac1 (rac1N17).
The membrane was visualised using a membrane tethered red fluorescent protein (ph-RFP). Each frame was obtained by combining a z-stack (composed of 15 optical sections separated by 1 µm) acquired …
Figure 2—video 3
Time lapse showing the dynamics of Sca::GFSTF (in green) in a SOP protrusion visualised using a membrane tethered RFP (in red).
Each frame was obtained by combining a z-stack (composed of three optical sections separated by 1 µm) acquired every 15 s. Time in min:s. Yellow arrowhead indicates a Sca vesicle traveling along the …
Figure 3 with 3 supplements
The mitotic wave of SOP cells is controlled by Notch/Delta/Scabrous signals.
(A) Heatmap of a representative heminota in control and scaBP2 homozygous null mutant aligned, anterior to the left. The relative time of SOPs division is colour coded according to the scale. (B) …
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Figure 3—source data 1
Related to Figure 3 Raw data.
- https://cdn.elifesciences.org/articles/75746/elife-75746-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
The fate of sensory organ cells is not affected in sca mutant.
Mechanosensory organ cells at two magnifications in control (A and C) and scaBP2 null mutant (B and D) pupae. Cells from sensory organs are identified by the neur> RFP construction (red). Fate of …
Figure 3—figure supplement 2
The amplitude of the mitotic wave is reduced in sca loss of function.
(A) The relative time of SOP cell division in control and scaBP2 homozygous mutant in row 1 (R1), row 2 (R2) and row 3 (R3) is plotted according to their relative position (mean time± SEM, of 16 …
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Figure 3—figure supplement 2—source data 1
Related to Figure 3—figure supplement 2 Raw data.
- https://cdn.elifesciences.org/articles/75746/elife-75746-fig3-figsupp2-data1-v2.xlsx
Figure 3—figure supplement 3
The best fit of the mitotic wave with a reduced amplitude is obtained after preferential lowering the inhibitory parameter μ.
The time of SOP cell division (mean time± SEM) in row 1 (R1), row 2 (R2) and row 3 (R3) is plotted relative to the position and time of division of the first cell to divide in each row. The …
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Figure 3—figure supplement 3—source data 1
Related to Figure 3—figure supplement 3 Raw data.
- https://cdn.elifesciences.org/articles/75746/elife-75746-fig3-figsupp3-data1-v2.xlsx
Figure 4 with 1 supplement
Microchæte axonal projections are affected after SOP mitotic wave disruption.
(A) Axon immunostaining using anti-Futsch/22C10 antibodies in control (w1118) and scaBP2 heminota at 24 hr APF aligned, anterior to the left. Axons in row one were artificially coloured in green for …
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Figure 4—source data 1
Related to Figure 4 Raw data.
- https://cdn.elifesciences.org/articles/75746/elife-75746-fig4-data1-v2.xlsx
Figure 4—figure supplement 1
sca downregulation modify the sensory neuropile structure.
On the left, the neuropile formed by all terminal axons from sensory organs on the central notum illustrating the absence (top) and presence (bottom) of the medial commissure (arrow) in thoracic …
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Figure 4—figure supplement 1—source data 1
Related to Figure 4—figure supplement 1 Raw data.
- https://cdn.elifesciences.org/articles/75746/elife-75746-fig4-figsupp1-data1-v2.xlsx
Figure 5 with 1 supplement
Disruption of the SOP mitotic wave leads to changes in fly behaviour.
(A) Eight frames of a time-lapse recording of a decapitated control fly showing the sequence of movements of the metathoracic leg (artificially coloured in green) upon stimulation of the dorsal …
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Figure 5—source data 1
Related to Figure 5 Raw data.
- https://cdn.elifesciences.org/articles/75746/elife-75746-fig5-data1-v2.xlsx
Figure 5—video 1
Recording of cleaning reflex assay in a decapitated control adult fly.
The leg movements observed were elicited after stimulation by air puffs of sensory organs in the more posterior central part of the notum. Air puffs were materialised by a red dot.
Figure 6
A neural progenitor mitotic wave is required for asynchronous axon outgrowth.
The mitotic wave of precursor cells induces an asynchronous arrival of sensory axons into the thoracic ganglion. This is depicted here by axons coloured according to the developmental time of …
Appendix 1—figure 1
Example of the phenomenological model with and for .
For for low to strong inhibition. Please note that the y scales are different.
Appendix 1—figure 2
TOP LEFT: Simulation different production rate (other parameters are , , , ).
TOP RIGHT: Simulation with various temporal constant (other parameters are, , ,, ). This adjusts the temporal scale but not the shape of the wave BOTTOM LEFT simulation with various …
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Appendix 1—figure 2—source code 1
Source code for model description.
- https://cdn.elifesciences.org/articles/75746/elife-75746-app1-fig2-code1-v2.zip
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Appendix 1—figure 2—source code 2
Source code for minimal model description.
- https://cdn.elifesciences.org/articles/75746/elife-75746-app1-fig2-code2-v2.zip
Videos
Video 1
Time lapse recording of a control pupa during a 4 hr period beginning at 15 hr after pupal formation.
The wave of SOP division is highlight in red in the row 1. Each frame was obtained by combining a z-stack (composed of 10 optical sections separated by 2 µm) acquired every 3 min. During the in vivo …
Video 2
Time lapse recording of the SOP mitotic wave in control pupa (top panel) and in scaBP2 mutant pupa (bottom panel).
For comparison, two hemi-nota are aligned. The observed difference in SOP nuclei size in the two movies is due to the lineage markers used (nuclear GFP and histone-RFP). The waves of SOP divisions …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Antibody | Anti-Futsch (clone 22C10, mouse monoclonal) | DSHB | AB_528403 | IF (1:200) |
| Antibody | Anti-HA (clone 3F10, rabbit polyclonal) | Roche | AB_231 4,622 | IF (1:250) |
| Antibody | Anti-Flag (M2, mouse monoclonal) | Sigma | AB_259529 | IF (1:250) |
| Antibody | Anti-V5 (E10/V4RR), DyLight650 (mouse monoclonal) | ThermoFisher | AB_2537642 | IF (1:100) |
| Antibody | Anti-cut (mouse monoclonal) | DSHB | AB_528186 | IF (1:500) |
| Antibody | Anti-ELAV (rat monoclonal) | DSHB | AB_7E8A10 | IF (1:500) |
| Antibody | Anti-Su(H) (rat polyclonal) | Gift F. Schweisguth | IF (1:500) | |
| Antibody | Anti-Prospero (mouse monoclonal) | DSHB | AB_528440 | IF (1:500) |
| Antibody | Anti-pdm (rabbit polyclonal) | Gift T. Preat | IF (1:2000) | |
| Antibody | Anti-Dl (mouse monoclonal) | DSHB | AB_528194 | IF (1: 500) |
| Antibody | Anti-GFP (B-2, rabbit polyclonal) | Santa Cruz | sc-9996 | IF (1: 500) |
| Antibody | Goat anti-rabbit AlexaFluor 488 | Molecular Probes | AB_2576217 | IF (1: 1000) |
| Antibody | Goat anti-rat AlexaFluor 488 | Molecular Probes | AB_2534074 | IF (1: 1000) |
| Antibody | Goat anti-mouse AlexaFluor 488 | Molecular Probes | AB_2534088 | IF (1: 1000) |
| Antibody | Goat anti-rat AlexaFluor 568 | Molecular Probes | AB_2534121 | IF (1: 1000) |
| Antibody | Goat anti-mouse AlexaFluor 568 | Molecular Probes | AB_144696 | IF (1: 1000) |
| genetic reagent (D. melanogaster) | w1118 | Bloomington Stock Center | BDSC_3605 | |
| genetic reagent (D. melanogaster) | neurD> GFP | F.Schweisguth | ||
| genetic reagent (D. melanogaster) | scaBP2 | Bloomington Stock Center | BDSC_7320 | |
| genetic reagent (D. melanogaster) | UAS sca-RNAi | Vienna Drosophila Resource Center | VDRC#44,527 | |
| genetic reagent (D. melanogaster) | sca::GFSTF | Bloomington Stock Center | BDSC_64443 | |
| genetic reagent (D. melanogaster) | neur> ph (PLCd)::RFP | F.Schweisguth | ||
| genetic reagent (D. melanogaster) | UAS-MCFO-1 | Bloomington Stock Center | BDSC_64085 | |
| genetic reagent (D. melanogaster) | UAS-rac1N17 | Bloomington Stock Center | BDSC_6292 | |
| genetic reagent (D. melanogaster) | Dl7 | Bloomington Stock Center | BDSC_485 | |
| genetic reagent (D. melanogaster) | UAS-ph::GFP | Y.Bellaiche | ||
| genetic reagent (D. melanogaster) | neurp72> Gal4 | Y.Bellaiche |
Additional files
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Transparent reporting form
- https://cdn.elifesciences.org/articles/75746/elife-75746-transrepform1-v2.docx
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Appendix 1—figure 2—source code 1
Source code for model description.
- https://cdn.elifesciences.org/articles/75746/elife-75746-app1-fig2-code1-v2.zip
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Appendix 1—figure 2—source code 2
Source code for minimal model description.
- https://cdn.elifesciences.org/articles/75746/elife-75746-app1-fig2-code2-v2.zip
