Pattern regulation in a regenerating jellyfish
- Sorbonne Université, CNRS, Laboratoire de Biologie du Développement de Villefranche-sur-mer (LBDV), France
Figures
Figure 1 with 2 supplements
Regenerative potential of Clytia hemisphaerica jellyfish.
(A–D) Anatomy of Clytia jellyfish. (A) Swimming female jellyfish (m: manubrium; g: gonads; tb: tentacle bulbs). (B) Tetraradial body organization: each umbrella quadrant comprises a radial canal …
Figure 1—figure supplement 1
Organ regeneration in relation to feeding status.
(A,B) Examples of organ ablation in fed individuals, each illustrated by a cartoon showing the cut (as an orange dotted line) and images of the regenerate at successive times. The regenerated gonad …
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Figure 1—figure supplement 1—source data 1
Raw data for the plots shown in Figure 1—figure supplement 1C–G.
- https://cdn.elifesciences.org/articles/54868/elife-54868-fig1-figsupp1-data1-v2.xlsx
Figure 1—figure supplement 2
Umbrella restorative potential.
Examples of different kinds of ablation. Left cartoon illustrates the cut (as an orange dotted line, removed parts are veiled), in the middle is an image of the regenerate at 24 hpd, and to the …
Figure 2 with 1 supplement
Manubrium regeneration, but not umbrella remodeling, requires cell proliferation.
(A) Top: schematics of the different stages of manubrium regeneration following manubrium ablation. Below: Phalloidin staining (white) and EdU incorporation (1 hr pulse) at successive time points …
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Figure 2—source data 1
Raw data for the plots shown in Figure 2B,C and Figure 2—figure supplement 1B,E.
- https://cdn.elifesciences.org/articles/54868/elife-54868-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
Cell proliferation increases in the radial canals during manubrium regeneration, and is efficiently blocked by hydroxyurea treatment.
(A, B) EdU incorporation at successive time points following manubrium ablation (1 hr pulse) detects cell proliferation in the MG-segment of radial canals. (A) Images of EdU detection (red in top …
Figure 3 with 1 supplement
Dynamics of i-cell migration during manubrium regeneration.
(A) Cartoon illustrates the distribution of CheNanos1-expressing cells (in red), also shown by FISH images in the bottom panels (CheNanos1+ cells in red; nuclei counter-stained with Hoechst, in …
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Figure 3—source data 1
Raw data for the plots shown in Figure 3H.
- https://cdn.elifesciences.org/articles/54868/elife-54868-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
I-cell migration during manubrium regeneration.
(A) Fluorescent in situ hybridization (FISH) showing the localization of CheNanos1-expressing cells (shown in red), in a two-week-old non-operated medusa. Blue: Hoechst-counter-stained nuclei, here …
Figure 4
Non-targeted cell recruitment of Mobilizing Gastro-Digestive (MGD) cells during regeneration.
(A) Images illustrating the distribution of MGD cells in Clytia organs (manubrium, gonads, tentacle bulbs), as indicated for each row. MGD cells can be recognized by the orange/brown pigment (DIC …
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Figure 4—source data 1
Raw data for the plots shown in Figure 4D.
- https://cdn.elifesciences.org/articles/54868/elife-54868-fig4-data1-v2.xlsx
Figure 5
Radial canals locally dictate the morphology of regenerating manubrium.
Cartoons illustrate experimental designs, with orange dotted lines showing cuts; fluorescence images show nuclear Hoechst staining (in white) of regenerated manubria (6 dpd), with blue dashed lines …
Figure 6 with 1 supplement
Smooth muscle hubs mark the position of regenerating manubria.
For each experiment, cartoons illustrate the cut (orange dotted lines), and stereomicroscope images the responses of live regenerates, while fluorescent images show phalloidin staining (in white) of …
Figure 6—figure supplement 1
Multiple muscle hubs correlate with the regeneration of multiple manubria.
For each experiment, cartoons illustrate the cut (orange dotted lines), and stereomicroscope images the responses of live regenerates, while fluorescent images show phalloidin staining (in white) of …
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Figure 6—figure supplement 1—source data 1
Raw data for the plots shown in Figure 6—figure supplement 1D.
- https://cdn.elifesciences.org/articles/54868/elife-54868-fig6-figsupp1-data1-v2.xlsx
Figure 7
Effects of additional manubria on manubrium regeneration.
Cartoons illustrate the grafting experiments, showing the ablation (orange dotted lines) and the grafted manubria position (in magenta). In (A–E), stereomicroscope images show live regenerates, in (F…
Figure 8 with 1 supplement
Actomyosin-driven subumbrella remodeling restores medusa shape.
(A) Morphology of remodeling bisected medusae; successive stereomicroscope images showing the rapid reduction of exposed edges. After closure of the wound, the manubrium migrates towards the center, …
Figure 8—figure supplement 1
Actomyosin-driven subumbrella remodeling restores medusa shape.
(A) Anti phospho-Myosin Light Chain 2 (Ser19) antibody (here indicated as PMyo) staining (upper row) of intact and remodeling muscle fibers, visualized by phalloidin staining (bottom row). PMyo …
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Figure 8—figure supplement 1—source data 1
Raw data for the plots shown in Figure 8—figure supplement 1C.
- https://cdn.elifesciences.org/articles/54868/elife-54868-fig8-figsupp1-data1-v2.xlsx
Figure 9 with 1 supplement
Hub stabilization is required for manubrium blastema formation.
(A,B) Successive images (0, 8, 10, 24 hpd and 4 dpd) of remodeling jellyfish fragments, deriving from an asymmetric cut across the umbrella, which generates one smaller fragment (‘S’) without …
Figure 9—figure supplement 1
Muscle hub dynamics in proximal manubrium grafts.
Dynamics of smooth muscle fibers in the case of grafted and manubrium-ablated jellyfish (cartoon). Representative images are shown for each case: 12 hpd most (n: 6/8) specimens show an organized hub …
Figure 10 with 2 supplements
Wnt/ß-catenin signaling drives manubrium regeneration.
(A) Sampling strategy for transcriptomic analyses: fragments of the central umbrella (orange dotted area) were collected at different times, prior (n.d.) or following manubrium ablation. (B) Heatmap …
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Figure 10—source data 1
Raw expression data used to construct the heatmap shown in Figure 10B; information about the RNAseq samples; expression values (tpm) for all the Clytia hemisphaerica transcripts for the 12 RNAseq samples.
- https://cdn.elifesciences.org/articles/54868/elife-54868-fig10-data1-v2.xlsx
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Figure 10—source data 2
Primers and raw data of the qPCR experiments shown in Figures 10E and 11C and Figure 11—figure supplement 1.
- https://cdn.elifesciences.org/articles/54868/elife-54868-fig10-data2-v2.xlsx
Figure 10—figure supplement 1
PCA plot of RNAseq samples from the manubrium area.
Non-dissected (n.d.), 0 hpd, 6 hpd, 12 hpd, 24 hpd (hub stabilization), 48 hpd (blastema onset). Two biological replicates of pooled samples were sequenced, per condition. PCA demonstrates samples …
Figure 10—figure supplement 2
PKF118-310 treatment of embryos.
DIC images of Clytia embryos incubated from the 4-cell stage in 0.7-µM PKF118-310 (middle and right) or in FASW (left). The spherical morphology and lack of endoderm of drug-treated embryos at 24 …
Figure 11 with 1 supplement
Remodeling-dependent activation of CheWnt6 expression.
(A) Schematics of S/L fragments to the left, and stereomicroscope/in situ hybridization images of 8 hpd control and blebbistatin-treated to the right. At 8 hpd CheWnt6 expression was detected at the …
Figure 11—figure supplement 1
Treatment with blebbistatin inhibits early increase of CheWnt6 expression at wound area.
(A) Schematic of ‘f’ and ‘F’ dissected tissue fragments from: (i) non-dissected (n.d.) and non-treated controls, (ii) non-dissected, blebbistatin-treated for 8 hr, (iii) control S fragments 8 hpd …
Figure 12
Summary figure.
Actomyosin-driven remodeling restores the circular shape (phase i), and generates a transient hub of remodeling smooth muscle fibers expressing CheWnt6. If stabilized after the re-centering step (pha…
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Clytia hemisphaerica) | CheNanos1 | GenBank | JQ397274 | |
| Gene (Clytia hemisphaerica) | CheWnt3 | GenBank | EU374721 | |
| Gene (Clytia hemisphaerica) | CheWnt6 | GenBank | EU374719 | Named CheWnt6 following Condamine et al., 2019 |
| Gene (Clytia hemisphaerica) | CheAxin | GenBank | EU374716 | |
| Gene (Clytia hemisphaerica) | Cheβ-cat | GenBank | JQ438997 | |
| Gene (Clytia hemisphaerica) | CheBra1 | GenBank | DQ872898 | |
| Gene (Clytia hemisphaerica) | CheFoxA | GenBank | GBGP01000196 | |
| Gene (Clytia hemisphaerica) | Z4B strain | Leclère et al., 2019 | Polyp colony producing female jellyfish | |
| Antibody | Rhodamine goat anti-rabbit secondary antibody | Jackson ImmunoResearch | IF (1:500) | |
| Antibody | Alexa Fluor 488 or 594 goat anti-rat secondary antibodies | Thermo Fisher Scientific | #A-11006 #A-11007 | IF (1:200) |
| Antibody | anti-Myosin Light Chain two phosphorylated at serine19 (PMyo) rabbit polyclonal | Cell signaling technology | #3671 | IF (1:200) |
| Antibody | anti-tyrosinated tubulin antibody (YL1/2) rat monoclonal | Abcam | #6160 | IF (1:50) |
| Chemical compound, drug | Hydroxyurea | Sigma-Aldrich | #H8627 | |
| Chemical compound, drug | Blebbistatin | Sigma-Aldrich | #B0560 | |
| Chemical compound, drug | BDM (2,3-Butanedione monoxime) | Sigma-Aldrich | #B0753 | |
| Chemical compound, drug | Menthol | Sigma-Aldrich | #M2772 | |
| Chemical compound, drug | PKF118-310 | Sigma-Aldrich | #K4394 | |
| Commercial assay, kit | EdU Click-It kit (Alexa Fluor 555) | Thermo Fisher Scientific | #C10338 | |
| Other | FluoSpheres | Molecular Probes | #F8812 | |
| Other | DiI | Thermo Fisher Scientific | #D3911 | |
| Other | Alexa Fluor 488 Phalloidin | Thermo Fisher Scientific | #10125092 | (1:100) |
| Other | Hoechst dye | Sigma-Aldrich | #94403 | (1 µg/ml) |
| Other | Reference transcriptome (Clytia hemisphaerica) | Leclère et al., 2019 | ENA identifier: PRJEB28006 | Genome guided transcriptome |
| Other | RNAseq samples (Clytia hemisphaerica) | This study | ENA identifier: PRJEB37920 | BGISEQ-500 50nt single reads generated for 12 RNA samples:pooled samples of umbrella central area collected at six time points during manubrium regeneration (two biological replicates for each). |
