Figure 1.
Tomo-seq allows the identification of specific gene expression patterns restricted to the (ventral) aortic microenvironment. (A-D) Schemes of the embryo species (top panels) and samples (bottom panels) used for tomo-seq analyses. E3 chicken anterior slices (A), 35-day-old human slices (B), E10.5 and E11.5 mouse slices (C), 28- and 40-hpf whole zebrafish embryos (D, left panel), and 28- and 40-hpf zebrafish trunks (D, right panel). The red areas mark the (ventral) aortic region where cluster and HSC emergence occurs. (E-G) Immunostainings of chicken (E), human (F), and mouse (G) embryo slices showing the location of clusters in the aorta. (E) Thick transverse slice of an E3 chicken embryo (slice adjacent to the one used for tomo-seq) stained with anti-MEP21 (red), anti-CD45 (green), and anti-RUNX1 (blue) antibodies. (F) Aorta of human embryo slice 1 stained with anti-CD34 (red), anti-CD45 (green), and anti-RUNX1 (white) antibodies and 4′,6-diamidino-2-phenylindole (blue). (G) Aorta of an E10.5, thick mouse embryo slice stained with anti-CD31 (red) and anti-KIT (green) antibodies. Arrows, dorsal clusters. (H) Global view of a Tg(kdrl:mCherry;cd41:eGFP) reporter zebrafish embryo at 40 hpf; mCherry fluorescence highlights the whole vasculature, GFP fluorescence highlights HSPCs. (I) Trunk region containing the aortic region and yolk sac elongation of the 40-hpf zebrafish embryo shown in panel H (dashed boxed). Single fluorescent pictures (right panels). (J-L) Thick slices from E3 chicken (J), 35 day-old human (K), and E11.5 mouse (L) embryos used for tomo-seq. (M) Whole 40-hpf zebrafish embryo used for tomo-seq. (N) Dissected trunk region of a 40-hpf zebrafish embryo used for tomo-seq. (O) Tomo-seq traces for MPL (red) and SHH (black) along the dorsal-ventral axis of the E3 chicken embryo slice shown in panel J. Colored areas corresponding to the ventral part of the aorta (R1, red) or to the dorsal part of the aorta (R2, blue). The numbers on the x-axes of z-score plots represent sequential individual section numbers (in O,R-S,V,X). (P-Q) ISH for MPL (P) and SHH (Q) on transverse sections of E3 chicken embryo, showing the restricted expression of MPL and SHH to clusters and notochord, respectively. Dashed boxed areas are enlarged in the right panels. (R) Tomo-seq traces for BMP4 (blue), GATA2 (red), and WWP2 (black) along the dorsal-ventral axis of human embryo slice 1, shown in panel K. Colored areas corresponding to the ventral part of the aorta (R1, red) or to the dorsal part of the aorta (R2, blue). (S) Tomo-seq traces for Gata2 (red), Bmp4 (blue), and Shh (black), along the dorsal-ventral axis of the E11.5 mouse embryo slice shown in panel L. Colored areas corresponding to the aortic region (R1, red) or to the regions flanking the aorta (R2 and R3, blue). (T-U) ISH for Bmp4 (T) and Gata2 (U) on E11.5 mouse embryo transverse sections. Dashed boxed areas are enlarged in the right panels. (V) Tomo-seq traces for pax6a (blue), nkx2.1a (green), hoxc11a (black), and apoa1a (red) along the posterior-anterior axis of the 40-hpf zebrafish embryo shown in panel M. (W) WISH showing the expression patterns of pax6a, nkx2.1a, and hoxc11a in 40-hpf zebrafish embryos. (X) Tomo-seq traces for apoa1a (blue), shha (black), and cxcl12b (red) along the dorsal-ventral axis of the 40-hpf zebrafish embryo trunk shown in N. Red area (R1): slices corresponding to the yolk sac elongation region (including the aorta) (V) or the aortic region (X). Blue areas (R2 and R3): slices corresponding to regions flanking the aorta region. (Y) WISH showing the expression patterns of apoa1a, shha, and cxcl12b in 40-hpf zebrafish embryos (left panels). Dashed boxed areas (trunk regions) are enlarged in the middle panels. Embryos were cryosectioned post-ISH to show the expression pattern of apoa1a, shha, and cxcl12b along the dorsal-ventral axis of the embryo trunks (right panels). The total number of embryos analyzed for WISH is indicated on the pictures. A, anterior; Ao, aorta; D, dorsal; Nc, notochord; NT, neural tube; Me, mesonephros; P, posterior; R, region; V, ventral; YS, yolk sac; YSe, yolk sac elongation. Bars represent 50 μm (E-G,Y [right panels]), 100 μm (J,P-Q), 150 μm (L,T-U), and 200 μm (H,K,M,W,Y [left panels]).

Tomo-seq allows the identification of specific gene expression patterns restricted to the (ventral) aortic microenvironment. (A-D) Schemes of the embryo species (top panels) and samples (bottom panels) used for tomo-seq analyses. E3 chicken anterior slices (A), 35-day-old human slices (B), E10.5 and E11.5 mouse slices (C), 28- and 40-hpf whole zebrafish embryos (D, left panel), and 28- and 40-hpf zebrafish trunks (D, right panel). The red areas mark the (ventral) aortic region where cluster and HSC emergence occurs. (E-G) Immunostainings of chicken (E), human (F), and mouse (G) embryo slices showing the location of clusters in the aorta. (E) Thick transverse slice of an E3 chicken embryo (slice adjacent to the one used for tomo-seq) stained with anti-MEP21 (red), anti-CD45 (green), and anti-RUNX1 (blue) antibodies. (F) Aorta of human embryo slice 1 stained with anti-CD34 (red), anti-CD45 (green), and anti-RUNX1 (white) antibodies and 4′,6-diamidino-2-phenylindole (blue). (G) Aorta of an E10.5, thick mouse embryo slice stained with anti-CD31 (red) and anti-KIT (green) antibodies. Arrows, dorsal clusters. (H) Global view of a Tg(kdrl:mCherry;cd41:eGFP) reporter zebrafish embryo at 40 hpf; mCherry fluorescence highlights the whole vasculature, GFP fluorescence highlights HSPCs. (I) Trunk region containing the aortic region and yolk sac elongation of the 40-hpf zebrafish embryo shown in panel H (dashed boxed). Single fluorescent pictures (right panels). (J-L) Thick slices from E3 chicken (J), 35 day-old human (K), and E11.5 mouse (L) embryos used for tomo-seq. (M) Whole 40-hpf zebrafish embryo used for tomo-seq. (N) Dissected trunk region of a 40-hpf zebrafish embryo used for tomo-seq. (O) Tomo-seq traces for MPL (red) and SHH (black) along the dorsal-ventral axis of the E3 chicken embryo slice shown in panel J. Colored areas corresponding to the ventral part of the aorta (R1, red) or to the dorsal part of the aorta (R2, blue). The numbers on the x-axes of z-score plots represent sequential individual section numbers (in O,R-S,V,X). (P-Q) ISH for MPL (P) and SHH (Q) on transverse sections of E3 chicken embryo, showing the restricted expression of MPL and SHH to clusters and notochord, respectively. Dashed boxed areas are enlarged in the right panels. (R) Tomo-seq traces for BMP4 (blue), GATA2 (red), and WWP2 (black) along the dorsal-ventral axis of human embryo slice 1, shown in panel K. Colored areas corresponding to the ventral part of the aorta (R1, red) or to the dorsal part of the aorta (R2, blue). (S) Tomo-seq traces for Gata2 (red), Bmp4 (blue), and Shh (black), along the dorsal-ventral axis of the E11.5 mouse embryo slice shown in panel L. Colored areas corresponding to the aortic region (R1, red) or to the regions flanking the aorta (R2 and R3, blue). (T-U) ISH for Bmp4 (T) and Gata2 (U) on E11.5 mouse embryo transverse sections. Dashed boxed areas are enlarged in the right panels. (V) Tomo-seq traces for pax6a (blue), nkx2.1a (green), hoxc11a (black), and apoa1a (red) along the posterior-anterior axis of the 40-hpf zebrafish embryo shown in panel M. (W) WISH showing the expression patterns of pax6a, nkx2.1a, and hoxc11a in 40-hpf zebrafish embryos. (X) Tomo-seq traces for apoa1a (blue), shha (black), and cxcl12b (red) along the dorsal-ventral axis of the 40-hpf zebrafish embryo trunk shown in N. Red area (R1): slices corresponding to the yolk sac elongation region (including the aorta) (V) or the aortic region (X). Blue areas (R2 and R3): slices corresponding to regions flanking the aorta region. (Y) WISH showing the expression patterns of apoa1a, shha, and cxcl12b in 40-hpf zebrafish embryos (left panels). Dashed boxed areas (trunk regions) are enlarged in the middle panels. Embryos were cryosectioned post-ISH to show the expression pattern of apoa1a, shha, and cxcl12b along the dorsal-ventral axis of the embryo trunks (right panels). The total number of embryos analyzed for WISH is indicated on the pictures. A, anterior; Ao, aorta; D, dorsal; Nc, notochord; NT, neural tube; Me, mesonephros; P, posterior; R, region; V, ventral; YS, yolk sac; YSe, yolk sac elongation. Bars represent 50 μm (E-G,Y [right panels]), 100 μm (J,P-Q), 150 μm (L,T-U), and 200 μm (H,K,M,W,Y [left panels]).

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal