The Glide/Gcm fate determinant controls initiation of collective cell migration by regulating Frazzled
- Institut de Génétique et de Biologie Moléculaire et Cellulaire, France
- Centre National de la Recherche Scientifique, UMR7104, France
- Institut National de la Santé et de la Recherche Médicale, U964, France
- Université de Strasbourg, France
- Tata Institute for Fundamental Research, India
Figures
Figure 1 with 1 supplement
Expression of Fra in wing glia.
(a–c) Immunolabeled repo>GFP wing (glia in green: anti-GFP; neurons in red: anti-22c10) at different stages. (a) Initiation of migration, (b) reaching the level of the costa, and (c) migration …
Figure 1—figure supplement 1
Expression profile of Fra.
(a–f) 18 hAPF repo>GFP/+ wing, immunolabeled with anti-22c10 (neurons in red), anti-Fra (gray) and anti-GFP (glia in green) shows the presence of Fra in the glial soma (white arrowheads) at the …
Figure 2
Role of Fra in wing glia.
(a, b) 28 hAPF wings labeled with anti-Repo (glial nuclei): (a) WT wing showing complete migration;(b) fra3/+ wing showing incomplete migration (dashed box). (c) Histogram representing the migratory …
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Figure 2—source data 1
Migratory index and repo count of of fra3 wings in WT background.
This file contains underlying raw data for Figure 2.
- https://doi.org/10.7554/eLife.15983.005
Figure 3 with 2 supplements
An instructive role of the chemoattractant receptor Fra in collective glia migration.
(a–b) Histogram representing the MI upon fra knock down (fra KD) or overexpression (fra GOF) using the gcm>GFP/+ line. In (b), the MI was calculated at 24 hAPF. The MI was calculated using the …
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Figure 3—source data 1
Migratory index and time-lapse analysis of fra conditional mutants in the gcm>GFP/+ background.
This file contains underlying raw data for Figure 3.
- https://doi.org/10.7554/eLife.15983.007
Figure 3—figure supplement 1
Migratory and neuronal defects in the frazzled mutation.
(a) Histogram representing the MI of the indicated genotypes, calculated using the membrane GFP transgenic line (UAS-mCD8-GFP). (b) Histogram showing the percentage of wings with axonal delay in gcm>…
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Figure 3—figure supplement 1—source data 1
Summarizing the role of neurons in glia migration and the genetic interaction between gcm and fra in different transheterozygote combinations.
This file contains underlying source data for Figure 3—figure supplement 1 and 2.
- https://doi.org/10.7554/eLife.15983.009
Figure 3—figure supplement 2
Speed analysis and genetic interaction between gcm and fra in glia migration.
(a) Graph showing the speed of the most-proximal glial cell of the chain (μm/h, y-axis; hAPF, x-axis) in gcm>GFP/+ and in gcm>fra GOF glia. The distance covered by the front cell is measured by …
Figure 4
Gcm affects collective glia migration in a dose-dependent manner.
(a) MI in fra mutants, gcm LOF and rescues of the indicated genotypes: gcm>GFP/+, gcm/fra LOF, gcm>gcm LOF, gcm>gcm KD and gcm>gcm GOF wings. (b) MI calculated at 24 hAPF in gcm>gcm GOF wings. The …
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Figure 4—source data 1
Summary of the role of gcm in glia migration.
This file contains underlying raw data for Figure 4.
- https://doi.org/10.7554/eLife.15983.012
Figure 5 with 1 supplement
The effects of Gcm on collective glia migration are independent of the number of Repo-positive nuclei.
(a) Bar chart showing the numbers of L1 glial nuclei in the various genotypes. (b) MI in the various genotypes, calculated using the membrane GFP transgenic line.
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Figure 5—source data 1
Effects of gcm on glia migration are independent of any change in the number of repo+ cells.
This file contains underlying raw data for Figure 5.
- https://doi.org/10.7554/eLife.15983.014
Figure 5—figure supplement 1
Fate conversion and Gcm expression in blood cells do not explain the gcm migratory phenotype.
(a–a”) gcm>GFP/+ and (b–b”) gcm>GFP/+;UAS-gcm-RNAi/+ 17 hAPF wings immunolabeled with anti-Elav (neuronal nuclei in blue) and anti-Repo (glial nuclei in red). Note the glial cells converted into …
Figure 6 with 1 supplement
fra is a direct Gcm target.
(a) Schematic representation of the fra gene indicated in blue. Lines indicate the introns, thin regions indicate the untranslated sequences, thick regions indicate the coding exons, pale blue …
Figure 6—figure supplement 1
Assays confirming fra as a direct Gcm target.
(a) Western blot analysis on total protein extracts of transfected S2 cells showing the GFP levels from the fra-GFP reporter plasmid (representative Western Blot shows one out of a sample of three). …
Figure 7 with 2 supplements
NetB serves as a chemoattractant in collective glia migration.
(a) MI of the indicated genotypes. Histogram shows the MIs quantified for WT, for NetAΔ or NetBΔ single mutant wings and for NetAΔNetBTM or NetAΔNetBmyc wings. (b) NP4151-Gal4 driven GFP expression …
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Figure 7—source data 1
Summary of the role of Netrins in glia migration.
This file contains underlying raw data for Figure 7.
- https://doi.org/10.7554/eLife.15983.019
Figure 7—figure supplement 1
Role of Netrins in collective glia migration.
(a–c) Graphs representing the MI of the indicated genotypes, calculated by nuclear labeling.
Figure 7—figure supplement 2
Early expression of NetB.
(a) Schematic of axonal bundles in a 10 hAPF wing, the green rectangle indicates the region shown in (b). (b) NP4151-Gal4-driven GFP expression of NetB in a 10 hAPF wing. Proximal NetB expression as …
Figure 8 with 1 supplement
Unc5 may act as a repellant in glia migration.
(a–d”’) Wing immunolabeled with anti-22c10 (red), anti-Unc5 (gray) and anti-GFP (green) in the transgenic line gcm>GFP/+ at different migratory stages. Unc5 starts being expressed in glia and …
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Figure 8—source data 1
Summary of role of unc5 in glia migration.
This file contains underlying raw data for Figure 8.
- https://doi.org/10.7554/eLife.15983.023
Figure 8—figure supplement 1
Unc5 in collective glia migration.
(a–a”) gcm>GFP/+ wing immunolabeled at 15 hAPF shows that the Unc5 protein cannot be detected at this stage. Note that (a–a”) are comprised of a few sections rather than maximum confocal …
Figure 9 with 1 supplement
Repo regulates Fra at late stages.
(a) Histogram showing the endogenous expression of fra upon S2 cell transfection with a Repo expression vector. The y-axis represents the relative expression levels in cells transfected with Repo …
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Figure 9—source data 1
Summary of role of repo in glia migration.
This file contains underlying raw data for Figure 9.
- https://doi.org/10.7554/eLife.15983.026
Figure 9—figure supplement 1
In vivo analysis of Unc5 GOF wings and fra KD wings using a late driver.
(a) Graphical representation of the migratory behavior of gcm>GFP/+ and gcm>unc5 GOF wings at the Initiation phase (n=9). (b) Box plot showing the raw data, the mean and the standard errors for the …
Author response image 1
Author response image 2
25 hAPF immunolabeled wing of the elav-Gal4,UASmCD8GFP transgenic line (anti-GFP in green and anti-Repo in red).
Note the presence of the mCD8GFP in the axons, in the glia (Repo-positive cells in red, arrows) and in the epithelium (asterisk).
Author response image 3
Author response image 4
Author response image 5
