Non-thalamic origin of zebrafish sensory nuclei implies convergent evolution of visual pathways in amniotes and teleosts
- Paris-Saclay Institute of Neuroscience (Neuro-PSI), Université Paris-Saclay, CNRS, France
- Laboratory of Fish Biology, Graduate School of Bioagricultural Sciences, Nagoya University, Japan
- Institute for Advanced Research, Nagoya University, Japan
- Laboratory of Molecular and Developmental Biology, National Institute of Genetics, Japan
- TEFOR Paris-Saclay, CNRS UMS2010, INRA UMS1451, Université Paris-Saclay, France
- Department of Genetics, SOKENDAI (The Graduate University for Advanced Studies), Japan
Figures
Figure 1
Tectofugal pathways in amniotes, amphibians, and teleosts and their phylogenetic relationships.
Schematic drawings of sagittal sections of the rodent, frog, and zebrafish brains are shown above a phylogenetic tree of bony vertebrates (Osteichthyes). Note that tetrapods (including mammals, …
Figure 2 with 2 supplements
GFP+ afferents in Tg(279A-GFP) zebrafish transgenic line.
(A and B) Frontal sections of Tg(279A-GFP) adult brain showing the GFP+ fibers in Dl of the pallium (A), and GFP+ cell bodies in PG (more specifically, PGl; B). The levels of the frontal sections …
Figure 2—figure supplement 1
GFP expression of the Tg(279A-GFP) adult brain.
(A and B) Sagittal sections of the Tg(279A-GFP) adult brain (anterior to the left). A is a more lateral section showing the PG and Dl, and B is a more medial section showing the PG axons coursing to …
Figure 2—figure supplement 2
Construction of Tg(gSAGFF279A) transgenic zebrafish.
(A) The structure of the gene trap construct T2KgSAGFF. (B) The integration site of T2KgSAGFF in the Tg(gSAGFF279A) line. The Tg(gSAGFF279A) fish was analyzed by southern blot, and DNA adjacent to …
Figure 3 with 1 supplement
Progression of GFP expression in the Tg(279A-GFP) brain during development.
Frontal sections of 4 wpf (A and B), 8 wpf (C and D), and the adult (E and F) brains at the level of the telencephalon (A, C, and E) and PG (B, D, and F). Approximate antero-posterior levels are …
Figure 3—figure supplement 1
GFP expression of the Tg(279A-GFP) brain at 4 wpf.
Frontal sections of the juvenile brain from anterior to posterior. Approximate levels of frontal sections are indicated in the schematic drawing on the top. The PGl-Dl projection is not observable …
Figure 4 with 1 supplement
Tract-tracing study showing connections of the lateral preglomerular nucleus (PGl).
Schematic drawings indicate the injection site of each tracer and levels of the frontal sections shown on the right panels. In all the frontal sections shown in A-L, the lateral side of the brain is …
Figure 4—figure supplement 1
Retinal projections terminating in TeO.
(A–D) Frontal sections of the adult TeO (lateral to the left) visualizing biocytin-labeled structures subsequent injection in the optic nerve. (A) The most anterior part of TeO shown with …
Figure 5 with 1 supplement
Progression of the mCherry+ cell distribution in the PG of Tg(her5:ERT2CreERT2;βactin:lox-stop-lox-hmgb1-mCherry) zebrafish treated with tamoxifen at 24hpf.
(A) In situ hybridization (ISH) of ert2Cre showing that the expression of Cre in this line is limited to the midbrain-hindbrain boundary (MHB) at 24 hpf. (B–G) Sagittal (B, D, F) and frontal (C, E, G…
Figure 5—figure supplement 1
Distribution of mCherry+ cells in the PG following tamoxifen induction at 24 hpf in Tg(her5:ERT2CreERT2;βactin:lox-stop-lox-hmgb1-mCherry).
A and B show frontal sections of juvenile brains, and C–E show of adult brains. The level of the frontal sections is indicated in the schematic drawing on the top. (A and B) mCherry+ cells at the …
Figure 6 with 1 supplement
Co-localization of GFP and mCherry in the adult PG cells of the quadruple transgenic line Tg(her5:ERT2CreERT2;βactin:lox-stop-lox-hmgb1-mCherry;279A-GFP) following the tamoxifen induction at 24 hpf.
(A) Schematic drawing of the adult zebrafish brain indicating the level of section through the PG shown in B-F. (B) A single plane confocal image showing a global view of the frontal section of a 3 …
Figure 6—figure supplement 1
A simplified schema of the constructs of transgenic lines and order of crossing to generate the first quadruple transgenic fish: Tg(her5:ERT2CreERT2;βactin:lox-stop-lox-hmgb1-mCherry;279A-GFP).
Figure 7 with 1 supplement
Expression of ert2Cre in the Tg(Dr830:ERT2CreERT2) embryonic brains.
(A–C) Sagittal sections of embryonic brains (rostral to the left). At 24 hpf (A), ert2Cre is highly expressed in the mesencephalic domain, and a weak expression is also found in the anterior part of …
Figure 7—figure supplement 1
Expression of ert2Cre in the Tg(Dr830:ERT2CreERT2) juvenile brains.
(A–F) Frontal sections of juvenile brains showing the telencephalic (Tel) and mesencephalic (Mes) areas. The levels of the frontal sections are indicated on the schematic drawing of a lateral view …
Figure 8 with 2 supplements
Co-localization of GFP and mCherry in the adult PG cells of the quadruple transgenic line Tg(Dr830:ERT2CreERT2;βactin:lox-stop-lox-hmgb1-mCherry;279A-GFP) following the tamoxifen induction at different developmental stages.
(A–G) Confocal images (5 µm projection) of frontal sections through the PG showing the co-localization of GFP and mCherry (arrowheads). The GFP/mCherry co-localization was consistently observed in …
Figure 8—figure supplement 1
A simplified schema of the constructs of transgenic lines and order of crossing to generate the second quadruple transgenic fish: Tg(Dr830:ERT2CreERT2;βactin:lox-stop-lox-hmgb1-mCherry;279A-GFP).
Figure 8—figure supplement 2
Proportion of mCherry+ cells during development in the PG of the quadruple transgenic line Tg(Dr830:ERT2CreERT2;βactin:lox-stop-lox-hmgb1-mCherry;279A-GFP).
Proportion of mCherry+ cells in relation to the total number of DAPI labelled cells was calculated. The dark grey represents the percentage of mCherry labeling observed after tamoxifen induction at …
Figure 9
Phylogenetic comparison of pallial projections in bony vertebrates (Osteichthyes).
Schematic drawing of representative embryonic brains of sarcopterygians (amniote and amphibian) and actinopterygians (teleost and Polypterus) are shown above the phylogenetic tree of the Osteichthyes…
Videos
Video 1
3D reconstruction of the PG-pallial visual afferents in the brain of the Tg(279A-GFP) transgenic line.
Confocal images of the entire zebrafish brain were reconstructed to demonstrate axonal projections in 3D. Selective visualization of GFP+ signal by manual segmentation (shown in orange) demonstrates …
Video 2
3D reconstruction of the mCherry+ cells in the 2 wpf brain of Tg(her5:ERT2CreERT2;βactin:lox-stop-lox-hmgb1-mCherry) zebrafish treated with tamoxifen at 24 hpf.
mCherry+ cells are shown in magenta, and DiD fiber labeling is shown in grey.
Tables
Appendix 1—key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Danio rerio) | Tg(gSAGFF279A;UAS:GFP) | This study | zTrap database: gSAGFF279A | transgenic zebrafish |
| Strain, strain background (Danio rerio) | Tg(UAS:GFP) | Asakawa and Kawakami, 2009 | zTrap database: UASGFP1A | transgenic zebrafish |
| Strain, strain background (Danio rerio) | Tg(her5:ERT2CreERT2) | Galant et al., 2016 | transgenic zebrafish | |
| Strain, strain background (Danio rerio) | Tg(Dr830:ERT2CreERT2) | Heuzé, 2017 | transgenic zebrafish | |
| Strain, strain background (Danio rerio) | Tg(βactin:lox-stop-lox-hmgb1-mCherry) | Wang et al., 2011 | transgenic zebrafish | |
| Antibody | chicken polyclonal anti-GFP | Aves Labs | Cat# GFP-1020; RRID:AB_10000240 | 1:1000 (brain sections), 1:400 (whole brain) |
| Antibody | rabbit polyclonal anti-dsRed | Takara Bio | Cat# 632496; RRID:AB_10013483 | 1:600 |
| Sequence-based reagent | oligonucleotide ert2 forward primer | Eurogentec (sequence based on Dirian et al., 2014) | 5'-ATGGCCGGTGACATGAGAGCTG-3' | |
| Sequence-based reagent | oligonucleotide Cre reverse primer | Eurogentec (sequence based on Dirian et al., 2014) | 5'-CATCAGGTTCTTCCTGACTTCAT-3' | |
| Chemical compound, drug | tamoxifen | Sigma-Aldrich | T176 | |
| Software, algorithm | Amira 6.5.0 | Thermo Fisher Scientific-FEI | ||
| Software, algorithm | Imaris 8.0.1 | Oxford Instruments | ||
| Software, algorithm | Fiji / ImageJ | Schindelin et al., 2012 | http://fiji.sc/ | |
| Software, algorithm | ImageJ cell counter plugin | Kurt De Vos (University of Sheffield) | https://imagej.nih.gov/ij/plugins/cell-counter.html | |
| Other | biocytin | Sigma-Aldrich | B4261 | |
| Other | biotinylated dextran amine (BDA) | Thermo Fisher Scientific | D7135 | |
| Other | DiI | Thermo Fisher Scientific | D7756 | |
| Other | DiD | Thermo Fisher Scientific | L7781 | |
| Other | DAPI | Sigma-Aldrich | 32670 |
Additional files
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Supplementary file 1
Conditions of tamoxifen treatments.
A summary table showing experimental conditions of tamoxifen treatments in two quadruple transgenic lines: Tg(her5:ERT2CreERT2;βactin:lox-stop-lox-hmgb1-mCherry;279A-GFP) and Tg(Dr830:ERT2CreERT2;βactin:lox-stop-lox-hmgb1-mCherry;279A-GFP). For the latter transgenic line, the conditions were adapted depending on the developmental stages.
- https://cdn.elifesciences.org/articles/54945/elife-54945-supp1-v1.xlsx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/54945/elife-54945-transrepform-v1.docx
