(A) Schematic representation of shape changes during the formation and looping of the heart tube (orange) in the E8.5 mouse embryo. Until E8.5f, the mouse embryo appears bilaterally symmetrical, and …
3D reconstructions of heart stages during the looping process, in a 3D pdf format.
3D visualisation of the heart, as reconstructed from HREM images, at the six stages shown in Figure 1. This file should be opened with Adobe Acrobat Reader. Click on the image to activate the manual rotation of the reconstruction. Each image may be rotated at will with the mouse (hold left click). Zoom in and out with the mouse wheel. Shortcuts at the top align all images in a ventral or dorsal view, with the notochord vertical.
(A) HREM image of an embryo section at E8.5f, with the notochord (green dot) and the centroid (red dot) of the myocardial tube (pale red) outlined. (B) Ventral view of a 3D reconstruction of the …
(A) Transverse section from an HREM image of an embryo at E8.5f showing the attachment of the heart tube to the body via the dorsal mesocardium (lateral thickness between red arrowheads). The …
(A) Transverse section from an HREM image of an embryo at E8.5f, in which the angles (right and left) between the heart tube and the dorsal pericardial wall, at the dorsal mesocardium, are shown. …
(A) Transverse embryo section at E8.5g, in which the width of the dorsal pericardial wall on either sides of the dorsal mesocardium is shown. The position of the section is indicated in brackets, as …
(A) 3D reconstruction of the heart tube at E8.5i, aligned with the notochord vertical (green), showing the axis of the myocardial tube (red) and the last tube section before the bifurcation (dotted …
(A) Timeline of the simulations, with the successive events reflecting experimental observations. (B) Simulation of heart shape changes in 3D with a Finite Element model of a straight tube, seen …
(A) Hollow-cylinder model of the heart tube, made of 1800 finite elements. (B) Mechanical constraints due to the attachment of the heart tube to the rest of the body: the arterial pole extremities …
In the computer simulation, parameters of pole asymmetries were explored to analyse the variety of output shapes of the heart tube. Simulated shapes are shown at step 80 with the ventral line in …
(A–B) Computer simulations of heart looping with the fate of the initial ventral line shown in blue. The simulated shape is shown at step 90 in ventral (left) and right-lateral (right) views. When …
Polr2aCreERT2/+; R26Rtdtomato/YFP embryos, are shown, after injection of a low dose of tamoxifen generating fluorescent cell mosaicism. (A–B) Initial and final images of a time-lapse movie between …
(A) Computer simulations of heart looping in control conditions (left), as in Figure 6, and when the dorsal mesocardium is persistent (right, simulated mutant). Simulated shapes are shown at step 90 …
3D reconstructions of all hearts of Shh-/- mutants and control littermates analysed at E8.5.
Ventral (left) and dorsal (right) views are shown, aligned with the notochord vertical (green). The myocardial layer (yellow) is made transparent, revealing the tube axis (red). Samples are identified with a number (S). The length of the heart tube is indicated, as well as the somite number (So) of the corresponding embryo. The genotype of control samples is Shh+/+ (S11, S17, S20) or Shh+/- (S8, S14). Scale bar: 100 µm.
(A, C) Lateral thickness of the dorsal mesocardium, measured as in Figure 3D, in five controls (A) and 5 Shh-/- mutant hearts (C) at E8.5. Samples are identified with a number (S), and the somite …
(A) 3D visualisation of HREM images of a control (left) and Shh-/- mutant (right) embryos, seen in a ventral view. The presence of the pericardium (S55) provides a rough appearance to the heart. (B, …
Comparison between quantitative predictions for four geometrical parameters raised by two types of computer simulations and biological measures in Shh-/-mutant hearts at E8.5. Simulation with 50% …
(A) Immunostaining of Mmp2, showing expression in the foregut (fg) at E8.5g, and vesicular localisations in the cardiac region (arrowheads in A1), but not in the neural tube (nt, A2). Maximum …
Video showing the successive sections of an E8.5h embryo, at the level of the heart. Images were acquired by HREM every 2 µm.
Video related to Figure 8—figure supplement 1A–C.
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Strain, strain background (Mus musculus) | wild-type, Swiss background | Janvier | ||
Strain, strain background (Mus musculus) | wild-type, C57Bl6 background | Janvier | ||
Strain, strain background (Mus musculus) | T4nLacZ, Swiss background | Biben et al. (1996) doi:10.1006/dbio.1996.0017 | PMID 8575622 | |
strain, strain background (Mus musculus) | Shh+/-, C57Bl6J background | Gonzalez-Reyes et al. (2012) doi:10.1016/j.neuron.2012.05.018 | MGI:5440762 | |
Strain, strain background (Mus musculus) | Polr2aCreERT2/+, C57Bl6 background | Guerra et al. (2003) PMID:12957286 | MGI:3772332 | |
Strain, strain background (Mus musculus) | R26YFP/+, C57Bl6 background | Srinivas et al. (2001) PMID:11299042 | MGI:2449038 | |
Strain, strain background (Mus musculus) | R26Rtdtomato/+ (Ai14), C57Bl6 background | Madisen et al. (2010) doi:10.1038/nn.2467 | MGI:3809524 | |
Antibody | anti-PH3 (rabbit monoclonal) | Abcam | Abcam: ab32107 | (1:100) |
Antibody | anti-Isl1 (mouse monoclonal) | Developmental Studies Hybridoma Bank | DSHB: 39.4D5 | (1:50) |
Antibody | anti-MMP2 (mouse monoclonal) | Santa Cruz | Santa Cuz: sc13594 | (1:50) |
Antibody | goat anti-rabbit IgG Alexa Fluor 546 | Invitrogen | Invitrogen: A11035 | (1:500) |
Antibody | goat anti-mouse IgG2b Alexa Fluor 488 | Invitrogen | Invitrogen: A21141 | (1:500) |
Antibody | goat anti-mouse IgG1 Alexa Fluor 488 | Invitrogen | Invitrogen: A21121 | (1:500) |
Commercial assay or kit | JB-4 embedding kit | Polysciences | Polysciences: 00226–1 | |
Chemical compound, drug | GM6001 (Ilomast) | Millipore | Millipore: CC1000 | 10 µM |
Software, algorithm | Gftbox | Kennaway et al. (2011) doi:10.1371/journal.pcbi.1002071 | Matlab Finite Element Analysis package simulating biological growth | |
Software | ICY | de Chaumont et al. 2012 doi:10.1038/nmeth.2075 | Open platform for bioimage informatics |
Code used to generate Figure 2F–G.
Code used to generate Figure 7.
Code used to generate Figure 8A.