Dpp dependent Hematopoietic stem cells give rise to Hh dependent blood progenitors in larval lymph gland of Drosophila
- Indian Institute of Science Education and Research Mohali, India
Figures
Figure 1 with 2 supplements
Presence of distinct cells in the first instar larval lymph glands of Drosophila that express several unique molecular markers.
(A) A schematic representation of lymph gland development throughout larval life. Red: progenitors; Magenta: niche cells; Blue: dorsal vessel (DV); orange: pericardial cells; light blue: …
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Figure 1—source data 1
Contains numerical data plotted in Figure 1Q and R and Total Fluorescence Intensity analyses for Q and R.
- https://doi.org/10.7554/eLife.18295.006
Figure 1—figure supplement 1
Notch expressing cells lack the earliest progenitor marker at first instar stage.
(A) dome-MESO-LacZ expression pattern at 3rd instar stage (n = 4) (-B–B") dome-MESO-LacZ (red; [B-B']) marks more number of progenitor cells in comparison to dome-Gal4, UAS-GFP (n = 8; green; [B–B"])…
Figure 1—figure supplement 2
Arrangement of Notch expressing cells with respect to rest of the cells in a first instar lymph gland.
(A–A') Anterior and lateral views of a 3D reconstruction image of a first instar lymph gland (lg) showing the position of N expressing cells (green) with respect to progenitor (red), niche (magenta) …
Figure 2 with 2 supplements
Cell cycle analysis of label retaining Notch expressing cells in first instar lymph gland of Drosophila.
(A–F) Transient activation of UAS-Histone2B::YFP by hs-Gal4 following scheme in (A) shows labeling of all cells of the embryonic lg at 2 hr post hs ([B–B""]; red; odd; n = 10). (C–F) Subsequent …
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Figure 2—source data 1
Contains numerical data plotted in Figure 2N and Figure 2—figure supplement 1E.
- https://doi.org/10.7554/eLife.18295.011
Figure 2—figure supplement 1
Cell cycle analyses of Notch expressing cells.
(A) Fluorescent ubiquitination based cell cycle indicator (Fucci), a fluorescent probe that labels cells in the S/G2/M phases (redrawn from Nakajima et al., 2011). Accumulation of GFP in the nucleus …
Figure 2—figure supplement 2
Absence of apoptosis in Notch expressing cells in first instar lymph gland of Drosophila.
(A–D") Time kinetics of N-Gal4, UAS-Apoliner show no apoptosis in N expressing cells at 8 hr (A–A"; n = 10), 13 hr (B–B"; n = 10), 18 hr (C–C"; n = 10) and 22 hr (D–D"; n = 10) AEH. Scale bar = 5 …
Figure 3 with 2 supplements
Notch expressing cells in first instar lymph gland of Drosophila are multipotent.
(A) N-GFP expression in a first instar lymph gland. N expression is detectable only till late first instar (also see Figure 1[E, M–O]). (B–E") shows that activation of G-TRACE with N Gal4, in a …
Figure 3—figure supplement 1
Fate map of the progenitors arising from Notch expressing HSCs.
(A) shows dome-GFP expression in progenitors of a late third instar lymph gland (n = 10). (B–C") show that dome expressing progenitors upon lineage tracing (C–C', green) from 18 hr AEH using the …
Figure 3—figure supplement 2
Notch requirement in crystal cell maintenance at a later stage in a third instar lymph gland.
(A–B") shows that, upon lineage tracing beyond HSC lifeline (following scheme in A), Notch lineage (green, [B, B"]) comprises of only crystal cells (red, live expression, [B, B'], n = 10). (C) Notch …
Figure 4
Notch is expressed earlier than Domeless in the embryonic lymph gland.
(A–B') Restriction of N-Gal4, UAS-2XEGFP expression in 4–5 cells (arrow heads) of late 16-stage embryonic lymph gland (Odd, red). A–A' shows lateral and B–B' depict dorsal views (n = 15). (C–F') …
Figure 5 with 1 supplement
Hierarchical relationship between dome and Notch expressing cells in the first instar lymph gland.
(A) Show the scheme of N lineage tracing followed throughout this panel. (B) Schematic representation of predicted cell types arising from N lineage tracing in A. A red cell indicates live …
Figure 5—figure supplement 1
Lineage tracing in an identical window reveals the hierarchical relationship between dome and Notch expressing cells.
(A–B) Expression of Ci (Cubitus interruptus, red: B–B"), a marker of progenitors in transient N lineage traced (green) lymph gland following the scheme in A (n = 10). (C) is a cartoon showing …
Figure 6 with 1 supplement
Genetic ablation of Notch expressing cells affects the size of the lymph gland.
(A) Expression of dome-MESO-LacZ in the lymph gland 16 hr after hatching. Four cells (arrow) lacking dome-MESO-LacZ expression are seen. (B) dome- MESO-LacZ expression in third instar larval lymph …
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Figure 6—source data 1
Contains numerical data plotted in Figure 6E,F.
- https://doi.org/10.7554/eLife.18295.020
Figure 6—figure supplement 1
HSCs and progenitor lineages in the lymph gland.
A scheme showing fate maps of progenitor and N expressing HSCs. (A) Progenitor cells give rise to all cells of the lymph gland. (B) N expressing cells are founder cells of progenitors, thus can also …
Figure 7 with 1 supplement
Requirement of Decapentapelagic (Dpp) from the PSC for hematopoietic stem cell maintenance in Drosophila.
(A) The maintenance signal for progenitors: Hedgehog (green; n = 7) starts to express in lg at about 18 hr AEH. (B–C') Lamin C (red) is expression in the niche/PSC (green) in 8 hr and 18 hr lg. (D–D"…
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Figure 7—source data 1
Contains numerical data plotted in Figure 7R and Figure 7—figure supplement 1D,M.
- https://doi.org/10.7554/eLife.18295.023
Figure 7—figure supplement 1
Involvement of Dpp signaling in HSC maintenance.
(A–B) show Upd3 expression ([A], green, n = 12) in a subset of PSC cells not earlier than 18 hr AEH, while (B) shows Wingless (Wg, red, n = 16) expression throughout first instar lymph gland. (C–C") …
Figure 8
Significant parallels in HSC development in Mammalian and Drosophila hematopoiesis.
Models depicting similarities in the development of HSCs from the mammalian AGM (A) and D. melanogaster’s cardiogenic mesoderm (B; redrawn from (Mandal et al., 2004). The cardiogenic mesoderm in Dros…
Author response image 1
Videos
Video 1
Position of Notch expressing cells in the first instar lymph gland with respect to progenitors (Red), PSC (Magenta) and the dorsal vessel (Blue).
Also see Figure 1—figure supplement 2.
