A molecular mechanism for the topographic alignment of convergent neural maps
- University of Strasbourg, France
- Cambridge Computational Biology Institute, University of Cambridge, United Kingdom
- University of Strasbourg Institute of Advanced Study, France
- The Salk Institute for Biological Studies, United States
Figures
Figure 1 with 1 supplement
Validation of the Efna3 KI model I.
(A–F) Immunochemical staining using anti-EFNA3 (LS-C6547, LS-Bio) in P1 (A, B) and P8 (C) WT (n = 3 animals) and P1 (D, E) and P8 (F) Efna3KI/KI (n = 5 animals) RGCs. In (E, F) arrows indicate EFNA3 …
Figure 1—figure supplement 1
Scheme of the knock-in strategy generating the Isl2-ires-Efna3 mutant mouse.
https://doi.org/10.7554/eLife.20470.003
Figure 2
Validation of the Efna3 KI model II.
(A, B) Synaptophysin (red) and DAPI (blue) staining in P34 WT (n = 4 animals) (A) and Efna3KI/KI (n = 4 animals) (B) retinas. Scale bars represent 50 µm. (C) Efna mRNAs relative to nasal WT …
Figure 3 with 2 supplements
Retinal anterograde mapping and retrograde DiI labelling in P8 animals.
(A) Retino-collicular maps generated by the Leave-One-Out method at P8 in WT (n = 38 animals), Efna3KI/+ (n = 15 animals) and Efna3KI/KI (n = 43 animals) mutants. Grey lines represent each map for …
Figure 3—figure supplement 1
Anterograde DiI labelling showing the focal injection in the retina (white star in A) and the corresponding termination zones in the SC (B).
Scale bars represent 400 µm. D, dorsal; N, nasal; T, temporal, V, ventral; R, rostral; C, caudal; SC, superior colliculus.
Figure 3—figure supplement 2
Representative examples of collicular parasagittal sections showing labelled anterograde retinal termination zones in P8 Efna3KI/KI (A) and WT (B) littermates.
Insets show the corresponding retinal DiI injection indicated by a white arrow. Scales bars represent 400 µm. N, nasal; T, temporal; R, rostral, C, caudal; SC, superior colliculus; WT, wild-type.
Figure 4 with 4 supplements
Anterograde cortico-collicular mapping in P14 animals.
(A–C) Cortico-collicular maps generated by the Leave-One-Out method at P15 in Efna3KI/KI (n = 19 animals) (A), Efna3KI/+ (n = 16 animals) (B) and WT (n = 9 animals) (C) mutant littermates. Grey …
Figure 4—figure supplement 1
Representative examples of collicular parasagittal sections showing cortical terminations zones and the corresponding injections in V1 in Efna3KI/KI (A), Efna3KI/+ (B) and WT (C) P15 littermates.
White panels show the termination zones analyses using ImageJ. Scale bars represent 1 mm (V1 whole-mount), 500 μm (SC), R: rostral, C: caudal, L: lateral, M: medial; WT, wild-type.
Figure 4—figure supplement 2
Collicular termination zones on parasagittal sections revealed after double DiI – DiD V1 anterograde injections in P15 Efna3KI/KI (A) and WT (B).
White panels show the corresponding ImageJ termination zones analyses. Scale bars represent 1 mm (V1) SC, 500 µm (SC). R, rostral; C, caudal; L, lateral; M, medial; SC, superior colliculus; WT, …
Figure 4—figure supplement 3
Immunostaining revealing EFNA3 expression (green) in P3 (A, D) and P8 (B–F) Efna3KI/KI and WT SC parasagittal sections after retinal anterograde DiI injection (red fibers).
Scale bars represent 30 µm (A, D), 10 μm (B - F).
Figure 4—figure supplement 4
EFNA3 expression in Efna3 mutants and WT optic nerves.
(A–F) Immunohistochemical staining revealing EFNA3 expression (green) in P3 and P8 Efna3KI/KI and WT SC parasagittal sections after retinal anterograde DiI injection (red fibers). Scale bars …
Figure 5 with 1 supplement
V1 retrograde labelling and Ephas/Efnas quantitative expression in V1 and SC.
(A) Retrograde labelling by collicular DiI injection in P14 Efna3KI/KI (n = 4 animals) and WT (n = 3 animals) showing the localization of the projecting V1 cortical neurons in layer V (DiI, red; …
Figure 5—figure supplement 1
EFNA3/DAPI immunohistochemical staining in P3 (A – A’, (C, C’) and P8 (B – B’, (D), (D’ E) Efna3KI/KI (A – B’), WT (C – D’) and Efna3-/- (E) SCs sagittal sections.
R, rostral; C, caudal. Scale bars represent 200 µm (A–D’), 100 µm (E).
Figure 6 with 1 supplement
Retino- and cortico-collicular maps in Epha3KI/Efna3KI double heterozygous and compound Epha3KI/Efna3KI::Epha4 knock-in/out mutants.
(A) Co-immunostaining of EFNA3 (LS-C6547, LS-Bio; green) and EPHA3 (red) in P4 acutely isolated Epha3KI/Efna3KI (n = 8 animals/15 retinas) RGCs. Arrowheads indicate EPHA3/EFNA3 co-expression in the …
Figure 6—figure supplement 1
Retino-collicular map in P7 Epha3KI/+ mutants (n = 11 animals).
T, temporal; N, nasal; R, rostral; C, caudal.
Figure 7
Retinal Efnas quantification.
(A) Section of P2 mouse retina (n = 6 retinas from three animals from different litters) hybridized with Efna5, Efna2 and Efna3 probes. Quantification was performed for 10 segments of the RGC layer …
Figure 8
Theoretical analysis and modelling.
(A–I) Retino- (A, D, G) and cortico- (C, F, I) collicular maps generated by the 3-step Map alignment model in WT (A, C), Efna3KI/KI (D, F) and Efna3KI/+ (G, I) after n = 20 runs and 107 …
Figure 9
Schematic representation of the map alignment mechanism.
RGCs axons in WT and Efna3 KI animals project to the SC during the first postnatal week and form the retino-collicular map through forward signaling activated by collicular EFNAs and fiber-fiber …
Author response image 1
Retino-collicular projections analysis at P8 in ephrin-A3KI/KI∷EphA4+/- (A) and ephrin-A3KI/KI∷EphA4-/- (B) double mutants showing single retino-collicular maps (left panels) and examples of anterograde tracings (right panels: whole-mount confocal SCs images showing the single TZs – white spot).
Scale bars: 200 µm. R, rostral; C, caudal.
Author response image 2
Immunohistochemical staining on P3 and P8 ephrin-A3KI/KI and WT SC parasagittal sections revealing ephrin-A3 expression in collicular cells (controls are without primary anti-ephrin-A3 antibody).
Scale bars: 200 µm (A – D), 30 µm (A’ – D’, A’’, B’’); R, rostral; C, caudal.
Author response image 3
Immunostaining revealing ephrin-A3 expression (green) in P3 and P8 ephrin- A3KI/KI and WT SC parasagittal sections after retinal anterograde DiI injection (red fibers).
Scale bars: 30 µm (A, D), 10 µm (B – F).
Author response image 4
P3 ephrin-A3KI/KI (A-C’, G, G’) and WT (D-F’) optic nerve sections, at the proximal, medial and distal level from the optic disc, immunostained for ephrin-A3 (in green), DAPI (in blue) after retinal DiI injections (in red).
A control experiment (without ephrin-A3 antibody) is shown on ephrin-A3KI/KI proximal optic nerve section. Scale bars: 20 µm (A-G’), 10 µm (high magnification panels in A’, B’, D’, E’). Arrows in …
Author response image 5
Immunohistochemistry on P8 Efna3-/- (A, B, C, G, H, I) and Efna3+/- (D, E, F, J, K, L) tissue sections using EFNA3 antibodies from LSBio (LSC6547) (A-F) and Invitrogen (36-7500) (G-L).
Scale bars, 100µm (A, C, D, F, G, I, J, L), 50 µm (B, E, H, K).
Additional files
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Source code 1
Leave-one-out ‘Leave-one-out’ script in R.
- https://doi.org/10.7554/eLife.20470.020
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Source code 2
3-step map alignment model 3-step map alignment script in MATLAB
- https://doi.org/10.7554/eLife.20470.021
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Supplementary file 1
Sequences of the primers used for genotyping and expression analyses.
- https://doi.org/10.7554/eLife.20470.022
