Transgenic quails reveal dynamic TCF/β-catenin signaling during avian embryonic development
- Australian Regenerative Medicine Institute (ARMI), Monash University, Australia
- Institut NeuroMyoGène (INMG), University Claude Bernard Lyon1, CNRS UMR, France
- Monash Micro Imaging, Monash University, Australia
Figures
Figure 1 with 3 supplements
Generation of the 12xTF-d2GFP and the 16xTF-VNP transgenic Japanese quail lines.
(A) Vectors used to generate the 12xTFd2GFP line and (B) the 16xTF-VNP line. (C) An E3.5 12xTFd2GFP embryo and (D) an E3 16xTF-VNP embryo cleared with the 3DISCO method, showing an overview of the …
Figure 1—figure supplement 1
Expression of the 16xTF-VNP in the neural tube and migrating neural crest.
(A–D) Transverse sections of an E2.5 chick embryo electroporated in the NT with a DNA mix containing a CAG-TagBFP construct as a marker for electroporated cells, and the 16xTF-VNP construct. The …
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Figure 1—figure supplement 1—source data 1
Source data file for the quantification shown in Figure 1—figure supplement 1, panel N.
- https://cdn.elifesciences.org/articles/72098/elife-72098-fig1-figsupp1-data1-v2.xlsx
Figure 1—figure supplement 2
Comparison of reporter fluorescence levels.
(A–C) Confocal stack of a dorsal view of an E2.5 chick embryo electroporated in the NT with a DNA mix containing a CAG-TagBFP construct as a marker for electroporated cells and the 12xTFd2GFP. (D–F) …
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Figure 1—figure supplement 2—source data 1
Source data file for the quantification shown in Figure 1—figure supplement 2, panel J.
- https://cdn.elifesciences.org/articles/72098/elife-72098-fig1-figsupp2-data1-v2.xlsx
Figure 1—figure supplement 3
TCF/β-catenin reporter expression in E4 embryos.
(A–D) An HH23 12xTFd2GFP embryo immunostained for GFP (green), neurofilament (blue), and MyHC (red), showing the TCF/β-catenin reporter in the pharyngeal arches, maxilla, AER, and the ventrolateral …
Figure 2 with 3 supplements
The 16xTF-VNP reporter is a sensitive reporter of the TCF/β-catenin signaling activity.
Transverse sections at the levels of the front limb (A, C) and the trunk (B,D) of E3 12xTF-d2GFP (A–B) and 16xTF-VNP (C–D) transgenic embryos stained for GFP or mVenus (green), Pax7 (blue), MyHC …
Figure 2—figure supplement 1
Transverse sections at the levels of the front limb and the trunk of E3 12xTFd2GFP and 16xTF-VNP transgenic embryos stained for GFP or mVenus, respectively (green), Pax7 (blue), MyHC (red), and DAPI (gray).
The image details the different channels for Figure 2. Scale bar 50 µm. S, somite; NT, neural tube; LB, limb bud; AER, apical ectodermal ridge.
Figure 2—figure supplement 2
TCF/β-catenin reporter dynamic activity in NC cells.
Transverse sections of E3 (HH19) 16xTF-VNP stained for mVenus (green; A,D), HNK1 (red; B,D), and DAPI (blue; C,D). The TCF/β-catenin reporter is significantly upregulated in emerging NC cells and is …
Figure 2—figure supplement 3
In situ hybridization of Wnt3a and Wnt1 mRNA in quail embryos.
In situ hybridization of Wnt1 (A,B) and Wnt3a (C,D) in E3.5 quail embryos. Both Wnt1 and Wnt3a mRNA transcripts are presented in the NT (A,B,D). Wnt3a mRNA transcript is also present in the AER (C). …
Figure 3
Dynamic TCF/β-catenin signaling activity in somites.
An optical section of an E2.5 embryo somite stained for Pax7 (A,D); blue mVenus (B,D); green, and MyHC (C,D); red reveals high and low reporter expression levels in epithelial cells of the DML, …
Figure 4 with 2 supplements
TCF/β-catenin reporter expression in HH12 embryos.
Whole-mount view of HH12 12xTFd2GFP (A–C) and 16xTF-VNP (D–F) embryos immunostained for GFP or mVenus (green) and Pax7 (Red). Both embryos present strong TCF/β-catenin reporter activity in migrating …
Figure 4—figure supplement 1
Whole-mount view of HH4 16xTF-VNP embryo immunostained for mVenus (green), Pax7 (red), and DAPI (blue).
The TCF/β-catenin reporter activity is strongly expressed in the posterior half of the gastrulating embryo. Scale bar 200 µm. PS, primitive streak.
Figure 4—figure supplement 2
Whole-mount view of HH11 16xTF-VNP embryo immunostained for mVenus (green) and Pax7 (red).
This embryo presents strong TCF/β-catenin reporter activity in migrating cephalic neural crest in the head area, pharyngeal arches, somites, the posterior neural tube, PSM and the tail bud area. …
Figure 5
TCF/β-catenin reporter expression pattern in transverse sections of a 14HH 16xTF-VNP embryo.
Transverse sections at the level of the cervical somites (A–D), somite I (E–H), and the anterior PSM (I–L) of a 14HH 16xTF-VNP embryo, immunostained for mVenus (green), Acetylated Tubulin (red), and …
Figure 6
TCF/β-catenin reporter activity in late developmental stages.
Native TCF/β-catenin signaling reporter activity in E9 12xTF-d2GFP (panels A,B,D,E) and E8 16xTF-VNP embryos (C,F). In the 12xTFd2GFP embryo the TCF/β-catenin reporter is strongly expressed in the …
Videos
Video 1
A 3D reconstruction of E3.5 12xTFd2GFP embryo cleared with the 3DISCO method.
The embryo is stained for GFP (green), Pax7 (blue), and MyHC (red).
Video 2
A 3D reconstruction of E3 16xTF-VNP embryo cleared with the 3DISCO method.
The embryo is stained for GFP (green), Pax7 (blue), and MyHC (red).
Video 3
A 9 h time-lapse confocal analysis of an E2.5 12xTFd2GFP embryo somite.
The movie shows a single Z-plane of the somite (10 μm) and focuses on two cells in the dorsal DML which increase the TCF/β-catenin reporter activity (magenta arrowheads). We also show a cell …
Video 4
A 9 hr time-lapse movie of an E2 16xTF-VNP embryo showing the growing hind limb bud.
Ectodermal cells, strongly expressing the TCF/β-catenin reporter, are seen as they migrate toward the AER region where they condensate.
The migrating NC and somitic cells are also highly visible.
