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Dpp dependent Hematopoietic stem cells give rise to Hh dependent blood progenitors in larval lymph gland of Drosophila

View Article: PubMed Central - PubMed

ABSTRACT

Drosophila hematopoiesis bears striking resemblance with that of vertebrates, both in the context of distinct phases and the signaling molecules. Even though, there has been no evidence of Hematopoietic stem cells (HSCs) in Drosophila, the larval lymph gland with its Hedgehog dependent progenitors served as an invertebrate model of progenitor biology. Employing lineage-tracing analyses, we have now identified Notch expressing HSCs in the first instar larval lymph gland. Our studies clearly establish the hierarchical relationship between Notch expressing HSCs and the previously described Domeless expressing progenitors. These HSCs require Decapentapelagic (Dpp) signal from the hematopoietic niche for their maintenance in an identical manner to vertebrate aorta-gonadal-mesonephros (AGM) HSCs. Thus, this study not only extends the conservation across these divergent taxa, but also provides a new model that can be exploited to gain better insight into the AGM related Hematopoietic stem cells (HSCs).

Doi:: http://dx.doi.org/10.7554/eLife.18295.001

No MeSH data available.


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 indicates that the cell is in the S/G2 phase while its distribution into the cytoplasm corresponds to the initiation of M phase. (B–D) At 8 hr AEH, Fucci (green) accumulates in nucleus of N expressing cells ([B]; red; E(spl); n = 31) indicating S/G2 phase of cell cycle (also see Figure 2H–H"). N expressing cells (red, E(spl)) undergo asynchronous division from around 13 hr AEH indicated by cytoplasmic GFP ([C]; Fucci, arrowhead, n = 31, p=8.18287E-09, two tailed unpaired Student’s t-test; also see Figure 2I–I"). (D) No Fucci signal is detectable at 18 hr AEH ([H]; n = 31; p=4.63838E-13, two tailed unpaired Student’s t-test; (also see Figure 2M). (E) Quantitative analyses of the data shown above. The numbers of N expressing cell at different cell cycle stages with respect to cytoplasmic and nuclear GFP (Fucci) are counted for each hour. Error bars=SE. (F–H) N expressing cells undergo mitotic division from 13 hr AEH. (F) shows no PH3 incorporation at 8 hr. (G) Cytoplasmic distribution of Fucci corresponding to PH3 positive cells, is detected at 13 hr AEH. (H) Although there is PH3 incorporation in the lymph gland but no Fucci signal can be detected at 18 hr AEH (anti-phospho-histone H3, red). Scale Bar is  μm . Genotypes are shown on top of corresponding panels. DAPI marks the nucleus. Hours after larval hatching are as indicated in each panel.DOI:http://dx.doi.org/10.7554/eLife.18295.010
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fig2s1: 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 indicates that the cell is in the S/G2 phase while its distribution into the cytoplasm corresponds to the initiation of M phase. (B–D) At 8 hr AEH, Fucci (green) accumulates in nucleus of N expressing cells ([B]; red; E(spl); n = 31) indicating S/G2 phase of cell cycle (also see Figure 2H–H"). N expressing cells (red, E(spl)) undergo asynchronous division from around 13 hr AEH indicated by cytoplasmic GFP ([C]; Fucci, arrowhead, n = 31, p=8.18287E-09, two tailed unpaired Student’s t-test; also see Figure 2I–I"). (D) No Fucci signal is detectable at 18 hr AEH ([H]; n = 31; p=4.63838E-13, two tailed unpaired Student’s t-test; (also see Figure 2M). (E) Quantitative analyses of the data shown above. The numbers of N expressing cell at different cell cycle stages with respect to cytoplasmic and nuclear GFP (Fucci) are counted for each hour. Error bars=SE. (F–H) N expressing cells undergo mitotic division from 13 hr AEH. (F) shows no PH3 incorporation at 8 hr. (G) Cytoplasmic distribution of Fucci corresponding to PH3 positive cells, is detected at 13 hr AEH. (H) Although there is PH3 incorporation in the lymph gland but no Fucci signal can be detected at 18 hr AEH (anti-phospho-histone H3, red). Scale Bar is  μm . Genotypes are shown on top of corresponding panels. DAPI marks the nucleus. Hours after larval hatching are as indicated in each panel.DOI:http://dx.doi.org/10.7554/eLife.18295.010

Mentions: 10.7554/eLife.18295.009Figure 2—source data 1.Contains numerical data plotted in Figure 2N and Figure 2—figure supplement 1E.


Dpp dependent Hematopoietic stem cells give rise to Hh dependent blood progenitors in larval lymph gland of Drosophila
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 indicates that the cell is in the S/G2 phase while its distribution into the cytoplasm corresponds to the initiation of M phase. (B–D) At 8 hr AEH, Fucci (green) accumulates in nucleus of N expressing cells ([B]; red; E(spl); n = 31) indicating S/G2 phase of cell cycle (also see Figure 2H–H"). N expressing cells (red, E(spl)) undergo asynchronous division from around 13 hr AEH indicated by cytoplasmic GFP ([C]; Fucci, arrowhead, n = 31, p=8.18287E-09, two tailed unpaired Student’s t-test; also see Figure 2I–I"). (D) No Fucci signal is detectable at 18 hr AEH ([H]; n = 31; p=4.63838E-13, two tailed unpaired Student’s t-test; (also see Figure 2M). (E) Quantitative analyses of the data shown above. The numbers of N expressing cell at different cell cycle stages with respect to cytoplasmic and nuclear GFP (Fucci) are counted for each hour. Error bars=SE. (F–H) N expressing cells undergo mitotic division from 13 hr AEH. (F) shows no PH3 incorporation at 8 hr. (G) Cytoplasmic distribution of Fucci corresponding to PH3 positive cells, is detected at 13 hr AEH. (H) Although there is PH3 incorporation in the lymph gland but no Fucci signal can be detected at 18 hr AEH (anti-phospho-histone H3, red). Scale Bar is  μm . Genotypes are shown on top of corresponding panels. DAPI marks the nucleus. Hours after larval hatching are as indicated in each panel.DOI:http://dx.doi.org/10.7554/eLife.18295.010
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fig2s1: 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 indicates that the cell is in the S/G2 phase while its distribution into the cytoplasm corresponds to the initiation of M phase. (B–D) At 8 hr AEH, Fucci (green) accumulates in nucleus of N expressing cells ([B]; red; E(spl); n = 31) indicating S/G2 phase of cell cycle (also see Figure 2H–H"). N expressing cells (red, E(spl)) undergo asynchronous division from around 13 hr AEH indicated by cytoplasmic GFP ([C]; Fucci, arrowhead, n = 31, p=8.18287E-09, two tailed unpaired Student’s t-test; also see Figure 2I–I"). (D) No Fucci signal is detectable at 18 hr AEH ([H]; n = 31; p=4.63838E-13, two tailed unpaired Student’s t-test; (also see Figure 2M). (E) Quantitative analyses of the data shown above. The numbers of N expressing cell at different cell cycle stages with respect to cytoplasmic and nuclear GFP (Fucci) are counted for each hour. Error bars=SE. (F–H) N expressing cells undergo mitotic division from 13 hr AEH. (F) shows no PH3 incorporation at 8 hr. (G) Cytoplasmic distribution of Fucci corresponding to PH3 positive cells, is detected at 13 hr AEH. (H) Although there is PH3 incorporation in the lymph gland but no Fucci signal can be detected at 18 hr AEH (anti-phospho-histone H3, red). Scale Bar is  μm . Genotypes are shown on top of corresponding panels. DAPI marks the nucleus. Hours after larval hatching are as indicated in each panel.DOI:http://dx.doi.org/10.7554/eLife.18295.010
Mentions: 10.7554/eLife.18295.009Figure 2—source data 1.Contains numerical data plotted in Figure 2N and Figure 2—figure supplement 1E.

View Article: PubMed Central - PubMed

ABSTRACT

Drosophila hematopoiesis bears striking resemblance with that of vertebrates, both in the context of distinct phases and the signaling molecules. Even though, there has been no evidence of Hematopoietic stem cells (HSCs) in Drosophila, the larval lymph gland with its Hedgehog dependent progenitors served as an invertebrate model of progenitor biology. Employing lineage-tracing analyses, we have now identified Notch expressing HSCs in the first instar larval lymph gland. Our studies clearly establish the hierarchical relationship between Notch expressing HSCs and the previously described Domeless expressing progenitors. These HSCs require Decapentapelagic (Dpp) signal from the hematopoietic niche for their maintenance in an identical manner to vertebrate aorta-gonadal-mesonephros (AGM) HSCs. Thus, this study not only extends the conservation across these divergent taxa, but also provides a new model that can be exploited to gain better insight into the AGM related Hematopoietic stem cells (HSCs).

Doi:: http://dx.doi.org/10.7554/eLife.18295.001

No MeSH data available.