Ecdysone steroid hormone remote controls intestinal stem cell fate decisions via the PPARγ-homolog Eip75B in Drosophila
- Institute of Genetics, Heinrich-Heine-University, Germany
Figures
Figure 1 with 1 supplement
The Ecdysone receptor in intestinal progenitors controls tissue homeostasis.
(A) Scheme of the adult Drosophila melanogaster gastrointestinal tract with cartoon depicting the midgut epithelial monolayer composed of intestinal stem cells (ISC), enteroblasts (EB), enterocytes …
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Figure 1—source data 1
Data from Figure 1.
Panel G,H experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig1-data1-v2.xlsx
Figure 1—figure supplement 1
The EcR is expressed in the adult Drosophila midgut.
(A) Expression analysis of Ecdysone-receptor splice variants with specific primer sets performed on cDNA transcribed from mRNA isolations of whole midgut dissections of adult MF. (B–D) Antibody …
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Figure 1—figure supplement 1—source data 1
Data from Figure 1—figure supplement 1.
Panel H experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig1-figsupp1-data1-v2.xlsx
Figure 2 with 1 supplement
Intracellular 20-Hydroxy-ecdysone levels control ecdysone response through the ecdysone importer.
(A–C) Determination of 20HE titers in ovaries and hemolymph of wild-type (A,B) and ovoD1 (B,C) adult VF and MF 48 hr after mating. (A,C) show fold-change increases over VF titer (dotted line at y = 1…
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Figure 2—source data 1
Data from Figure 2.
Panel A,B,C,J,O,P,Q experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig2-data1-v2.xlsx
Figure 2—figure supplement 1
20HE regulates physiological adaptations of fatty acid metabolism.
(A) Drosophila food tubes of w1118 control egg layings to determine concentrations of RH5849. Please note that there is no crawling L3-larvae and puparium formation in both RH5849 concentrations. …
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Figure 2—figure supplement 1—source data 1
Data from Figure 2—figure supplement 1.
Panel H,O experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig2-figsupp1-data1-v2.xlsx
Figure 3 with 2 supplements
Ecdysone induced protein 75B is upregulated upon mating and controls progenitor differentiation.
(A) Cartoon depicting EcR signaling cascade activating early ecdysteroid target genes. (B) Expression analysis of Ecdysone- and JH-signaling target genes including protein isoforms on cDNA …
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Figure 3—source data 1
Data from Figure 3.
Panel C,L,M experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Analysis of Eip75B-A MARCM clones.
(A–E’’) Representative images of MARCM clones of the indicated allele five days after clone induction (ACI) counterstained with Dlg-1 and Pros immunohistochemistry. Control clones lead to proper EC …
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Figure 3—figure supplement 1—source data 1
Data from Figure 3—figure supplement 1.
Panel D,E,F,G experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig3-figsupp1-data1-v2.xlsx
Figure 3—figure supplement 2
EB specific genetic manipulation of Eip75B using kluReDDM.
(A) Cartoon depicting kluReDDM-tracing. klu-Gal4 is active in EC-committed EB only and EC progeny is labelled with nuclear H2B::RFP (Reiff et al., 2019). (B–D’’) Representative images of MF controls …
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Figure 3—figure supplement 2—source data 1
Data from Figure 3—figure supplement 2.
Panel A,J,K,L experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig3-figsupp2-data1-v2.xlsx
Figure 4
The Eip75B/PPARγ agonist Pioglitazone acts through Eip75B to stimulate progenitor differentiation.
(A–D) Representative images of adult PMG after seven days of esgReDDM tracing of control MF (A,B) and >Eip75 B-RNAi MF (C,D) fed with DMSO as control (A,C, 2.5 µl/ml food) and Pioglitazone (B,D; …
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Figure 4—source data 1
Data from Figure 4.
Panel E,F experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig4-data1-v2.xlsx
Figure 5
Crosstalk between JH- and Ecdysone-signaling pathways controlling intestinal progenitor proliferation and differentiation.
(A–B) Images of adult PMG of control VF (A) and forced expression of > Kr h1 (B) traced for seven days with esgReDDM. (C–H) Images of adult PMG with forced expression of > Kr h1 with > Eip75 B-RNAi …
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Figure 5—source data 1
Data from Figure 5.
Panel I,J experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig5-data1-v2.xlsx
Figure 6 with 1 supplement
Ecdysone induced protein 75B promotes EB differentiation in a Notch tumor paradigm.
(A) Cartoon depicting cell fate changes upon N-LOF combined with esgReDDM coloring (Ohlstein and Spradling, 2006; Ohlstein and Spradling, 2007). (B) > N RNAi driven by esgReDDM leads to different …
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Figure 6—source data 1
Data from Figure 6.
Panel F,I,J experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig6-data1-v2.xlsx
Figure 6—figure supplement 1
Size distribution of different Notch-tumor classes.
(A) Quantification of ISC progeny encompassing ISC-like, EC and EE classified after tumor size (classes I to IV) in R5 PMG of esgReDDM > N RNAi (n = 10,10,10). Error bars are Standard Error of the …
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Figure 6—figure supplement 1—source data 1
Data from Figure 6—figure supplement 1.
Panel A experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig6-figsupp1-data1-v2.xlsx
Figure 7 with 1 supplement
Ecdysone signaling promotes EC-fate in a Notch tumor paradigm.
(A–E’’) Pharmacological and genetic manipulation of Ecdysone signaling. Adult PMG of MF with N-LOF (>N RNAi or > NDN) treated with MeOH as control (A–A’’) or RH5849 (B–B’’) to activate Ecdysone …
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Figure 7—source data 1
Data from Figure 7.
Panel F,G experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig7-data1-v2.xlsx
Figure 7—figure supplement 1
Analysis of Notch mutant MARCM clones and consequences of clonal EcR-activation.
(A–E’’) Representative images of MARCM clones for the Notch receptor (N55e11) in MF PMG four days after clone induction (ACI) counterstained with Dlg-1 and Pros antibodies. Control Notch clones …
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Figure 7—figure supplement 1—source data 1
Data from Figure 7—figure supplement 1.
Panel C,G experimental data.
- https://cdn.elifesciences.org/articles/55795/elife-55795-fig7-figsupp1-data1-v2.xlsx
Author response image 1
Author response image 2
Author response image 3
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (Drosophila melanogaster) | esgReDDM | Antonello et al., 2015a DOI: 10.15252/embj.201591517 | Figures 1–7; Figure 1—figure supplement 1; Figure 3—figure supplement 1 | |
| Genetic reagent (D. melanogaster) | esgReDDM > Eip75B-A | Rabinovich et al., 2016 DOI: 10.1016/j.cell.2015.11.047 | Figure 3; Figure 3—figure supplement 1 | |
| Genetic reagent (D. melanogaster) | esgReDDM > Eip75B-B | Rabinovich et al., 2016 DOI: 10.1016/j.cell.2015.11.047 | Figure 3; Figure 3—figure supplement 1 | |
| Genetic reagent (D. melanogaster) | esgReDDM > Eip75B-C | Rabinovich et al., 2016 DOI: 10.1016/j.cell.2015.11.047 | Figure 3; Figure 3—figure supplement 1 | |
| Genetic reagent (D. melanogaster) | Srebp > CD8::GFP | Reiff et al., 2015 DOI: 10.7554/eLife.06930 | Figure 2—figure supplement 1 | |
| Genetic reagent (D. melanogaster) | Mex > ts | Phillips and Thomas, 2006 DOI: 10.1242/jcs.02839 | Figure 2—figure supplement 1 | |
| Genetic reagent (D. melanogaster) | MARCM (FRT2A) | Lee and Luo, 1999 DOI: 10.1016/S0896-6273 (00)80701–1 | Figure 3—figure supplement 1 | |
| Genetic reagent (D. melanogaster) | Eip75A81-MARCM (FRT2A) | Rabinovich et al., 2016 DOI: 10.1016/j.cell.2015.11.047 | Figure 3—figure supplement 1 | |
| Genetic reagent (D. melanogaster) | kluReDDM | Reiff et al., 2019 DOI: 10.15252/embj.2018101346 | Figure 3—figure supplement 2 | |
| Genetic reagent (D. melanogaster) | N55e11-MARCM (FRT19A) | Guo and Ohlstein, 2015 DOI: 10.1126/science.aab0988 | Figure 7—figure supplement 1 | |
| Chemical compund, drug | RH5849 | DrEhrenstorfer | DRE-C16813000 | 340 µM final concentration |
| Chemical compund, drug | Pioglitazone | Sigma-Aldrich | Sigma-Aldrich 112529-15-4 | 14 µM final concentration |
