Multi-view light-sheet imaging and tracking with the MaMuT software reveals the cell lineage of a direct developing arthropod limb
- Humboldt-Universität zu Berlin, Germany
- Imagopole, Citech, Institut Pasteur, France
- Max Planck Institute of Molecular Cell Biology and Genetics, Germany
- Howard Hughes Medical Institute, United States
- Max Delbrück Center for Molecular Medicine, Germany
Figures
Figure 1 with 2 supplements
Reconstruction of Parhyale embryogenesis with multi-view LSFM (see also Figure 1—video 1 and 2).
(A) Transgenic Parhyale embryo with H2B-mRFPruby-labeled nuclei mounted with fluorescent beads (green dots) for multi-view reconstruction. The embryo was imaged from the indicated 5 views with 45˚ …
Figure 1—video 1
Imaging Parhyale embryogenesis with multi-view LSFM
Time-lapse recording of a transgenic embryo from the crustacean amphipod Parhyale hawaiensis labeled with the nuclear H2B-mRFPruby fluorescent marker. The embryo was recorded on a Zeiss Lightsheet …
Figure 1—video 2
Imaging Parhyale embryogenesis with multi-view LSFM
Left side of the same embryo shown in Figure 1—video 1 rendered with the same settings but without rotation. Anterior is to the left and dorsal to the top.
Figure 2
Grid architecture of the Parhyale germband.
(A–A’’) Rendering of a Parhyale embryo at the growing germband stage: (A) Right, (A’) ventral, and (A’’) left side. Color masks indicate the anterior head region (blue), the bilaterally symmetric …
Figure 3 with 1 supplement
Cell tracking and lineage reconstruction with MaMuT (see also Figure 3—figure supplement 1).
(A) Workflow for image data analysis with MaMuT. Raw views (colored boxes in Multi-view Dataset) are registered (overlapping boxes in Multi-view Registration) and, optionally, fused into a single …
Figure 3—figure supplement 1
MaMuT layout.
(A–C) The three tabs of the MaMuT control panel. (A) The Views tab is used to launch and control the different displays of the image data and annotations. (B) The Annotation tab is used to define …
Figure 4 with 2 supplements
Early compartmentalization of the Parhyale thoracic limb (see also Figure 4—figure supplements 1 and 2).
(A–E) Lateral views of a Parhyale embryo rendered at the indicated developmental stages shown in hours (h) after egg-lay (AEL). Yellow masks show the left T2 limb (limb#1). (F–J’) Tracked cells …
Figure 4—figure supplement 1
Lineage reconstruction of the Parhyale thoracic limb.
(A–E) Lateral views of the same Parhyale embryo shown in Figure 4. Yellow masks indicate the left T2 limb (limb#1). (F–J) Tracked cells contributing to the T2 limb color-coded by the DV column they …
Figure 4—figure supplement 2
Independent evidence for early compartmentalization of the Parhyale thoracic limb.
(A–D) Lateral views of another Parhyale embryo imaged on a Zeiss LSFM prototype instrument rendered at the indicated developmental stages shown in hours (h) after egg-lay (AEL). Yellow masks …
Figure 5 with 2 supplements
Stereotyped and variable cell behaviors in developing Parhyale thoracic limbs (see also Figure 5—figure supplement 1, Figure 5—source data 1, and Figure 5—video 1).
(A–C) Schematic representations of the T2 limb primordium at the 4-row-parasegment stage displaying the 34 founder cells as squares color-coded based on their relative birth times: (A) limb#1, (B) …
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Figure 5—source data 1
Relative birth times of founder cells in Parhyale thoracic limbs.
- https://doi.org/10.7554/eLife.34410.013
Figure 5—figure supplement 1
Reconstructed lineage tree of a Parhyale T2 limb.
Each track resembles one or two of the 34 founder cells of limb#1 color-coded by their compartmental identity: anterior-dorsal in dark green, anterior-ventral in dark magenta, posterior-dorsal in …
Figure 5—video 1
Animation of tracked cells forming the Parhyale second thoracic limb
All tracked cells contributing to the T2 limb#1 are displayed as spheres of uniform color. The movie starts from the early limb specification stage at about 3 days AEL and covers limb bud formation …
Figure 6 with 2 supplements
Differential cell proliferation rates in the Parhyale thoracic limb (see also Figure 6—figure supplements 1 and 2).
(A–D) Lateral views of the same Parhyale embryo shown in Figure 4. (E–H’) Tracked cells in limb#1 were color-coded by their average cell cycle length according to the scale (in hours) shown at the …
Figure 6—figure supplement 1
Digital clonal analysis in the Parhyale thoracic limb.
Digital clones for each one of the 34 founder cells of the T2 limb#1 visualized at 114 hr (h) after egg-lay (AEL). In each panel, the name of the founder cell is shown in the top left corner and its …
Figure 6—figure supplement 2
Alternative quantifications of cell proliferation rates in the Parhyale thoracic limb.
(A –E) Tracked cells making up the T2 limb#1 shown at the indicated hours (h) after egg-lay (AEL) and color-coded by their compartmental identity: Anterior-Dorsal in dark green, Anterior-Ventral in …
Figure 7
Lineage comparisons within and across Parhyale thoracic limbs (see also Figure 7—source data 1).
(A) Hierarchical clustering of the 34 founder cells in the Parhyale T2 limb based on a distance matrix computed from their average division patterns in limb#1 and limb#2. The cluster of E4c3-c7 and …
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Figure 7—source data 1
Lineage distances between founder cells in Parhyale thoracic limbs.
- https://doi.org/10.7554/eLife.34410.020
Figure 8
Oriented cell divisions in the Parhyale thoracic limb.
(A–E) Cells in the T2 limb#1 shown at the indicated hours (h) after egg-lay (AEL) color-coded by the orientation of mitotic divisions relative to the AP boundary (cyan line). The AP boundary is …
Figure 9 with 1 supplement
Elaboration of the Parhyale limb PD axis (see also Figure 9—figure supplement 1).
(A–F) Rendering of the T2 limb#1 at the indicated hours (h) after egg-lay (AEL). The cells contributing to the T2 primordium are shown in cyan in panel A. Magenta dots indicate the tracked cells …
Figure 9—figure supplement 1
Proximal-distal lineage separation in the growing Parhyale thoracic limb.
(A–E) Tracked cells contributing to the T2 limb#1 color-coded by their compartmental identity: Anterior-Dorsal (dark green), Anterior-Ventral (dark magenta), Posterior-Dorsal (light green), and …
Figure 10 with 1 supplement
Analysis of developmental regulatory genes corroborates cellular models of limb morphogenesis (see also Figure 10—figure supplement 1).
(A–F) Brightfield images of T2, T3 and T4 limbs from S16-S18 embryos (top row, 84–96 hr AEL) and S19 embryos (bottom row, 96–108 hr AEL) stained by in situ hybridization for Ph-dpp (left columns), Ph…
Figure 10—figure supplement 1
Expression of Ph-dpp, Ph-Doc and Ph-H15 during Parhyale limb bud formation.
(A–F) Brightfield images of S16-S18 embryos (top row, 84–96 hr AEL) and S19 embryos (middle row, 96–108 hr AEL) stained by in situ hybridization for Ph-dpp (left columns), Ph-Doc (middle columns) …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Parhyale hawaiensis) | Wild Type | PMID: 15986449 | ||
| Strain, strain background (P. hawaiensis) | PhHS>H2B-mRFPruby | This paper | ||
| Recombinant DNA reagent | pMi{3xP3>EGFP; PhHS>H2B-mRFPruby} | This paper | ||
| Software, algorithm | MaMuT | This paper | http://imagej.net/MaMuT | |
| Software, algorithm | SIMI°BioCell | PMID: 9133433 | http://simi.com/en/products/cell-research | |
| Gene (P. hawaiensis) | Ph-dpp | This paper | GenBank: KY696711 | |
| Gene (P. hawaiensis) | Ph-Doc | This paper | GenBank: KY696712 | |
| Gene (P. hawaiensis) | Ph-en2 | This paper | GenBank: KY696713 | |
| Gene (P. hawaiensis) | Ph-H15 | This paper |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.34410.026
