Glial insulin regulates cooperative or antagonistic Golden goal/Flamingo interactions during photoreceptor axon guidance
- Graduate School of Life Science and Technology, Tokyo Institute of Technology, Japan
- Center for Transdisciplinary Research, Niigata University, Japan
- Brain Research Institute, Niigata University, Japan
- Mathematical Neuroscience Unit, Institute for Frontier Science Initiative, Kanazawa University, Japan
- Laboratory of Developmental Neurobiology, Graduate School of Medical Sciences, Kanazawa University, Japan
Figures
Figure 1 with 1 supplement
R8-specific labeling of Gogo and Fmi.
(A) Schematics of the Drosophila visual system in the third instar larva and the adult. (B) Schematics of the phase-specific R8 targeting during development. (C–G) Gogo localization at the terminals …
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Figure 1—source data 1
Source data for the quantification in Figure 1C-H.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig1-data1-v2.xls
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Figure 1—source data 2
Source data for the quantification in Figure 1N.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig1-data2-v2.xlsx
Figure 1—figure supplement 1
gogo expression gradually declines during midpupal stages.
(A–E) gogo expression level in R8 cells were monitored by gogo-Gal4 sensFLP UAS-FsF-mCD8GFP. gogo-Gal4 was created by inserting Gal4 into the gogo intron locus using MiMIC system. Photoreceptor …
Figure 2 with 3 supplements
Gogo and Fmi regulates the growth cone dynamic.
(A–L) The medulla of control, R8-specific gogo loss-of-function mutations, and R8-specific fmi loss-of-function was analyzed. (A–C) The medulla of the third instar larvae (phase 1) was labeled with …
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Figure 2—source data 1
Source data for the quantification in Figure 2A-C.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig2-data1-v2.xlsx
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Figure 2—source data 2
Source data for the quantification in Figure 2D.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig2-data2-v2.xlsx
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Figure 2—source data 3
Source data for the quantification in Figure 2H.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig2-data3-v2.xlsx
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Figure 2—source data 4
Source data for the quantification in Figure 2L.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig2-data4-v2.xlsx
Figure 2—figure supplement 1
R cell-specific loss-of-funtion of Gogo and Fmi.
(A–F) R axons in adult medulla visualized with GFP (green) counterstained with 24B10 (red) and anti-N-cadherin (blue) in control (A and D) and gogo (B and E), fmi (C and F) mutants. gogo and fmi …
Figure 2—video 1
Filopodial dynamics of the control animal.
Live imaging of R8 photoreceptor growth cone filopodial dynamics from early to midpupal stage in the control animal. Yellow arrows indicate R8 filopodia extension at step 3.
Figure 2—video 2
Filopodial dynamics of gogo mutant.
Live imaging of R8 photoreceptor growth cone filopodial dynamics from early to midpupal stage in gogo mutant. Yellow arrows indicate R8 filopodia extension at step 3, while the white arrow indicates …
Figure 3 with 2 supplements
Gogo has dual functions, 'cooperative’ and ‘antagonistic’ toward Fmi.
(A–E) R8 axons in wild type (A), R8-specific knockdowns of gogo (B), fmi (C), and gogo, fmi double knockdowns (D) in phase 1 were visualized using R8-specific UAS-mCD8GFP (green) counterstained with …
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Figure 3—source data 1
Source data for the quantification in Figure 3E.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig3-data1-v2.xlsx
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Figure 3—source data 2
Source data for the quantification in Figure 3H.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig3-data2-v2.xlsx
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Figure 3—source data 3
Source data for the quantification in Figure 3L.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig3-data3-v2.xlsx
Figure 3—figure supplement 1
Gogo and Fmi functions are not redundant.
(A–H) R8 axons in medulla at phase 1 (A–D) and phase 2 (E–H) were visualized with GFP (green) counterstained by anti-N-cadherin (magenta in A-D, blue in E-H) and 24B10 (red in E-H) in control (A, E) …
Figure 3—figure supplement 2
Functional domain analysis of Gogo.
(A–H) Small deletions as illustrated above each image heterozygous with gogo null mutation were analyzed at phase 1. R8 axons were labeled with myr-Tomato (green) counterstained with anti-N-cadherin …
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Figure 3—figure supplement 2—source data 1
Source data for the quantification in Figure 3—figure supplement 2O.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig3-figsupp2-data1-v2.xlsx
Figure 4
Gogo localization in R8 changes depending on the expression level of Fmi.
(A–H) Localization of R8-specific Gogo-GFP (A–E) and Fmi-mCherry (D–H) in loss-of-function (heterozygous mutation with R8-specific RNAi) or overexpression backgrounds. R8 axons were labeled with …
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Figure 4—source data 1
Source data for the quantification in Figure 4D-E.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig4-data1-v2.xlsx
Figure 5 with 1 supplement
Dual function of Gogo controlled by the phosphorylation of YYD motif.
(A–F) gogo rescue experiments in a background of gogo[H1675]/gogo[D1600] during phase 1 (third instar larva). R8 axons were visualized with mCD8GFP (green), and columns were labeled with N-cadherin …
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Figure 5—source data 1
Source data for the quantification in Figure 5F.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig5-data1-v2.xlsx
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Figure 5—source data 2
Source data for the quantification in Figure 5L.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig5-data2-v2.xlsx
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Figure 5—source data 3
Source data for the quantification in Figure 5Q.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig5-data3-v2.xlsx
Figure 5—figure supplement 1
Gogo and Fmi cytoplasmic domain change its functional properties.
(A) R8-specific hts loss-of-function animals were generated by hts heterozygote with R8-specific RNAi. In phase 2 (APF24%), hts loss-of-function shows R8 axons bundling phenotype (arrows) due to the …
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Figure 5—figure supplement 1—source data 1
Source data for the quantification in Figure 5—figure supplement 1B.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig5-figsupp1-data1-v2.xlsx
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Figure 5—figure supplement 1—source data 2
Source data for the quantification in Figure 5—figure supplement 1E.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig5-figsupp1-data2-v2.xlsx
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Figure 5—figure supplement 1—source data 3
Source data for the quantification in Figure 5—figure supplement 1H.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig5-figsupp1-data3-v2.xlsx
Figure 6 with 1 supplement
Glial insulin switches the Gogo-Fmi function from ‘cooperative’ to ‘antagonistic'.
(A–C) The phenotype of R8-specific dinr loss-of-function (dinr heterozygotes with R8 cell-specific RNAi) at the third instar larvae and APF24% (phase 1 and 2) was analyzed using R8-specific mCD8GFP …
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Figure 6—source data 1
Source data for the quantification in Figure 6C.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig6-data1-v2.xlsx
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Figure 6—source data 2
Source data for the quantification in Figure 6M.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig6-data2-v2.xlsx
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Figure 6—source data 3
Source data for the quantification in Figure 6Q.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig6-data3-v2.xlsx
Figure 6—figure supplement 1
Related to Figure 6 dilp genes expression pattern in optic lobe.
(A–I) dilp gene expression was monitored by dilp-Gal4, UAS-mCD8GFP (green) and counterstained with anti-Repo (red) and anti-N-cadherin (blue). dilp1, 2, 3, five are strongly expressed in …
Figure 7 with 2 supplements
Glial Fmi and R8 Gogo/Fmi instruct R8 to recognize the columnar center.
(A) R8 axon terminals visualized with myr-tdTomato (red, white) and glial cells visualized with mCD8GFP (green) and counterstained with anti-N-cadherin (blue) in phase 1 (third instar larva). The …
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Figure 7—source data 1
Source data for the quantification in Figure 7E.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig7-data1-v2.xlsx
Figure 7—figure supplement 1
Gogo and Fmi interact with Fmi to regulate cytoskeletal reorganization.
(A) In the phase 1, R7 neurons that are known to be the first core members of the medulla column formation were labeled with 20C11FLP, GMR-FsF-Gal4, UAS-mCD8GFP (magenta), and R axons with 24B10 …
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Figure 7—figure supplement 1—source data 1
Source data for the quantification in Figure 7—figure supplement 1J.
- https://cdn.elifesciences.org/articles/66718/elife-66718-fig7-figsupp1-data1-v2.xlsx
Figure 7—figure supplement 2
Genetic interaction between gogo and fmi in mushroom body.
(A–A’) Structure of mushroom bodies (MBs) in adult. PED, CA, and CB denote peduncle, calyx, and cell bodies, respectively. Dashed line rectangles in (A’) indicate the anterior region and posterior …
Additional files
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Supplementary file 1
List of genotypes used.
- https://cdn.elifesciences.org/articles/66718/elife-66718-supp1-v2.docx
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Supplementary file 2
oligo DNAs used for generating and analyzing transgenic flies.
- https://cdn.elifesciences.org/articles/66718/elife-66718-supp2-v2.docx
