Dermomyotome-derived endothelial cells migrate to the dorsal aorta to support hematopoietic stem cell emergence
- Department of Cell and Developmental Biology, University of California, San Diego, United States
- Department of Cell Differentiation, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic v.v.i, Czech Republic
- Department of Pharmacy, University of Oslo, Norway
Figures
Figure 1 with 3 supplements
Cell-type-specific endothelial cell markers highlight cellular diversity within the vasculature.
(A) Uniform manifold approximation projection (UMAP) plots of scRNA-seq data of total endothelial lineage cells collected from TgBAC(etv2:Kaede)ci6, Tg(fli1:DsRed)um13; Tg(tp1:GFP)um14, and Tg(drl:H2…
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Figure 1—source data 1
Transcriptomes of all endothelial cell clusters, myeloid, and erythroid cells.
The transcriptomes were extracted and read from cells purified collected from TgBAC(etv2:Kaede)ci6, Tg(fli1:DsRed)um13; Tg(tp1:GFP)um14, and Tg(drl:H2B-dendra) embryos at 22–24 hpf.
- https://cdn.elifesciences.org/articles/58300/elife-58300-fig1-data1-v2.xlsx
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Figure 1—source data 2
Comparison between Genes expressed in EC clusters (e.g. SDECs) and gene annotation of the same anatomical structure (e.g., somite) based on annotation from AmiGO (Consortium, 2019).
Expression of overlapping genes was compared in the same cluster (e.g. SDECs) between 15 ss and 22 hpf and divided into DE genes that are upregulated (Up) on downregulated (Down). Each DE group was then annotated using AmiGO (Consortium, 2019).
- https://cdn.elifesciences.org/articles/58300/elife-58300-fig1-data2-v2.xlsx
Figure 1—figure supplement 1
Cluster identity was assigned based on known marker genes.
(A–F) Following unsupervised clustering of single-cell transcriptomes, cluster identity was given based on known marker genes within established tissue lineages. Selected marker genes and the eight …
Figure 1—figure supplement 2
Comparison of BVECs-I cluster genes to brain annotated genes validates cluster origin.
(A) Uniform manifold approximation projection (UMAP) plots of scRNA-seq data of total endothelial lineage cells collected from TgBAC(etv2:Kaede)ci6, Tg(fli1:DsRed)um13; Tg(tp1:GFP)um14, and Tg(drl:H2…
Figure 1—figure supplement 3
Comparison of KVEC Cluster genes to kidney annotated genes validates cluster origin.
(A) Uniform manifold approximation projection (UMAP) plots of scRNA-seq data of total endothelial lineage cells collected from TgBAC(etv2:Kaede)ci6, Tg(fli1:DsRed)um13; Tg(tp1:GFP)um14, and Tg(drl:H2…
Figure 2 with 1 supplement
Cellular diversity within the vasculature can be traced back to the tailbud stage.
(A) Uniform manifold approximation projection (UMAP) plots of scRNA-seq data of total endothelial lineage cells collected from TgBAC(etv2:Kaede)ci6, Tg(fli1:DsRed)um13; Tg(tp1:GFP)um14, and Tg(drl:H2…
Figure 2—figure supplement 1
Differentially expressed genes between early and late ECs in BVECs-I or KVECs clusters highlight an early commitment to EC fate.
(A–D) Comparison of expression patterns of EC populations from early TgBAC(etv2:Kaede)ci6 15 ss and later 22 hpf etv2:Kaede+ ECs in the 63 overlapping genes between the BVECs-I transcriptome data …
Figure 3
Rare SDECs emerge from trunk somites and migrate to the dorsal aorta.
(A–D) Tg(actb2:nls-Eos); Tg(fli1:eGFP)y1 embryos were collected at developmental stages ranging from 4 to 18 ss. (A) Newly developed posterior somite pairs were selected by setting a region of …
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Figure 3—source data 1
A table summarizing all converted somite pairs and SDECs found in Tg(actb2:nls-Eos); Tg(fli1:eGFP)y1 embryos that were included in the final SDECs quantification assay.
Embryo’ somites were converted at developmental stages ranging from 4 to 18 somite stage (Column C) and imaged at 32–36 dpf. The imaging date (Column A), sample number within a cohort (Column B), and the number of observed SDECs (Column D) used for the quantification were documented, and the quantification and presented graph were done in Prism 9 (GraphPad).
- https://cdn.elifesciences.org/articles/58300/elife-58300-fig3-data1-v2.xlsx
Figure 4 with 2 supplements
Endothelial cells emerge from the dermomyotome at 12 ss.
(A–E) Time-lapse imaging from a dorsal view of Tg(etv2.1:EGFP)zf372 embryos injected with mOrange:CAAX mRNA and imaged between 10 ss and 15 ss. (A) The expression of Etv2:GFP+ cells is visible along …
Figure 4—video 1
Time-lapse imaging of Tg(etv2.1:eGFP)zf372; Tg(phldb1:mCherry) embryo between 12 and 16 ss.
Lateral view of a transgenic embryo. LPM cells migrate from the left side under the somites. SDECs (Green, arrows) arise from the first and fifth somites (Red) and exit the somites to follow their …
Figure 4—video 2
Time-lapse imaging of Tg(etv2.1:eGFP)zf372; Tg(phldb1:mCherry) embryo between 12 and 16 ss.
Lateral view of a transgenic embryo. SDECs (Green, arrows) arise from the somites (S, Magenta). By this time, most of the LPM cells (arrowheads) have ingressed underneath the somites. S, somites; …
Figure 5 with 1 supplement
notch is required for the maintenance of a bipotent skeletal muscle progenitor population in the somite.
(A–F) Dorsal view of 12 ss control (A–C) and meox1 morphant embryos (D–F). Embryos were submitted to double fluorescent in situ hybridization for meox1 (green) and etv2 (red). In control and …
Figure 5—figure supplement 1
Bipotent muscle progenitor cells contain endothelial potential that can reach the dermomyotome compartment.
(A) Max projection of 12 hpf notch3-/- mutant embryos injected with meox1 morpholino shows broad endothelial potential within the somite compartment by the ectopic formation of double positive meox1 …
Figure 6 with 1 supplement
npas4l is required for the specification of SDECs.
(A–D) WISH for etv2 in 12 ss npas4l-/- (cloche) mutant and control embryos. (B) cloche mutant embryos show an absence of etv2 expression along the A-P axis of the embryo, compared to sibling control …
Figure 6—figure supplement 1
npas4l is required for the specification of SDECs.
(A–D) Tg(etv2.1:eGFP)zf372; cloche mutant and heterozygous embryos were injected with meox1 and mib morpholinos. (C,D) Cloche mutant embryos showed loss of Etv2:eGFP expression in the LPM and …
Figure 7
Wnt signaling is required for the regionalization of SDECs.
(A) FISH for meox1 (red) and antibody staining for a destabilized Wnt/TCF reporter line (green) show co-expression of GFP and meox1 within the somite. (B) Inhibition of Wnt signaling using the …
Figure 8 with 1 supplement
SDECs contribute to the dorsal aorta but do not generate HSPCs.
(A–C) Lineage tracing of SDECs using tbx6:Gal4; Tg(UAS-Cre); A2BD shows dsRed+ cells in the vasculature region at 48 hpf (arrowheads). (E–J) Using a vasculature-specific switch line TgBACkdrl:LOXP-Am…
Figure 8—figure supplement 1
Paraxial mesoderm does not generate HSPCs.
(A) kdrl:Cre; A2BD and (B) tbx6:Gal4; Tg(UAS-Cre); A2BD adult kidney marrow was analyzed by flow cytometry. Top row illustrates that the hematopoietic lineages of kdrl:Cre; A2BD originate from …
Figure 9
SDECs act as a vascular niche for hemogenic endothelium.
(A) Heatmap of genes differentially regulated between LPM-derived endothelial cells (LPMDEC sample is composed of pre-HSCs and HE clusters) and SDECs. (B,C) Zebrafish embryos were injected with an …
