Fizzy-Related dictates A cell cycle switch during organ repair and tissue growth responses in the Drosophila hindgut
- Duke University School of Medicine, United States
Figures
Figure 1 with 1 supplement
Injured hindgut pyloric cells replace lost genome content using two distinct responses.
(A) Schematic of pyloric development. (B) Experimental injury scheme (see Results and Materials and methods). Numbers 1 and 2 are referenced in the text. (C–F) Adult pylori. Anterior boundary marked …
Figure 1—figure supplement 1
Supporting data related to Figure 1.
(A–D’) Larval and adult pylori. Cell death (TUNEL, green) and nuclei (DAPI, white). (A–A’) Uninjured larva. (B–B’) 16 hr larval injury. (C–C’) Uninjured Adult. (D–D’) 24 hr adult injury. (E) …
Figure 2 with 1 supplement
Distinct cell cycle programs underlie the distinct pyloric injury responses.
(A–B) Schematic of clone and injury induction in larvae (A) and adults (B). (C–F’) Clones in adult pylori. Clones (LacZ, green) and nuclei (DAPI, magenta). Yellow box highlights the region shown in …
Figure 2—figure supplement 1
Supporting data related to Figure 2.
(A) Clonal induction system used (see Materials and methods). (B) Quantification of clones induced per animal. Data represent mean ± SEM, N ≥ 10 animals, at least two replicates. Unpaired, two-tail …
Figure 3 with 1 supplement
DEMISE, a novel tool for dual control of site-specific genetic ablation and transgene expression.
(A–A’’’) DEMISE method and components (see Results and Materials and methods). (B–D) Adult pylori. Transgene expression marked by UAS-NLS GFP (green), cell death (TUNEL, magenta), and nuclei (DAPI, …
Figure 3—figure supplement 1
Supporting data related to Figure 3.
(A, B, C, D) Schematic of DEMISE experiment conditions for corresponding wing disc images on the right. GFP expression (green), FLP expression (magenta), apoptosis (red). (A’-A’, B’-B’, C’-C’) …
Figure 4 with 1 supplement
fizzy-related is a regulator of injury-mediated cell cycle programming.
(A–B’) Expression of the fzrG0418 enhancer trap (LacZ expression, green or grey), nuclei (DAPI, magenta) in uninjured larval pylorus (A–A’) or adult pylorus (B–B’). Yellow dashed line marks pyloric …
Figure 4—figure supplement 1
Supporting data related to Figure 4.
(A–A’) Rectal papillae, nuclei (DAPI, magenta) and Moesin-GFP (green) in control (A) and fzr RNAi (A’). (B–B’) Pyloric nuclei staining (DAPI, white) of uninjured control (B) and fzr RNAi (B’). (C) …
Figure 5 with 1 supplement
Endocycles preserve intestinal architecture in response to sustained tissue growth signaling.
(A) Schematic and (B) experimental design of sustained growth signaling expression. Numbers 1–3 in (B) are referenced in the text. (C–F) Adult pyloric nuclei (DAPI, white) ±RasVV12 expression. (C) …
Figure 5—figure supplement 1
Supporting data related to Figure 5.
(A,B) Adult pylori, nuclei (DAPI, white). Distinct Z-sections of the hindgut positions shown in each image are illustrated by a red line in an inset in the bottom left, where a black circle …
Tables
Table 1
DEMISE combinations tested in the hindgut
https://doi.org/10.7554/eLife.38327.009| Flip-out reaper insertion | Genotype | [ry[+t7.2]=hsFLP]1 | [ry[+t7.2]=hsFLP]12 |
|---|---|---|---|
| UAS-rprFLPout 10–3 | pUAST-FRT-STOP-FRT-rpr/CyO #Insertion 10–3 | Leaky | Not leaky (used in paper) |
| UAS-rprFLPout 9–1 | pUAST-FRT-STOP-FRT-rpr/TM3Sb #Insertion 9–1 | Leaky | Not leaky |
| UAS-rprFLPout 5–1 | pUAST-FRT-STOP-FRT-rpr/TM3Sb #Insertion 5–1 | Leaky | |
| UAS-rprFLPout 4–1 | pUAST-FRT-STOP-FRT-rpr/TM3Sb #Insertion 4–1 | Not leaky | |
| UAS-rprFLPout 10–1 | pUAST-FRT-STOP-FRT-rpr/CyO #Insertion 10–1 | Leaky | Leaky |
| UAS-rprFLPout 10–2 | pUAST-FRT-STOP-FRT-rpr/TM3Sb #Insertion 10–2 | Leaky | Leaky |
Key resources table
| Reagent type (species) | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| gene (D. melanogaster) | fzr | NA | FBgn0262699 | |
| genetic reagent (D. melanogaster) | byn > Gal4 | Singer et al.,1996 | FBal0137290 | P{GawB}bynGal4 |
| genetic reagent (D. melanogaster) | act >> LacZ | BDSC | Stock 6355 | P{ry[+t7.2]=Act5C (FRT.polyA)lacZ.nls1}3, ry[506] |
| genetic reagent (D. melanogaster) | hsFLP12;Sco/CyO | BDSC | Stock 1929 | P{ry[+t7.2]=hsFLP} 12, y[1] w[*]; sna[Sco]/CyO |
| genetic reagent (D. melanogaster) | UAS fzr RNAi | VDRC | Stock 25550 | w1118; P{GD9960}v25550 |
| genetic reagent (D. melanogaster) | UAS-Ras85DV12 | BDSC | Stock 4847 | w[1118]; P{w[+mC]=UAS-Ras85D.V12}TL1 |
| genetic reagent (D. melanogaster) | UAS-hid, UAS-reaper | Zhou et al 1997 | ||
| antibody | anti-Fasciclin III | DSHB | 7G10 | |
| antibody | anti-Phospho-Histone H3 | Cell Signaling | ab9706 | |
| antibody | anti-BrdU | Serotec | 3J9 | |
| antibody | anti-Beta-Galactosidase | Abcam | ab9361 | |
| recombinant DNA reagent | pUAST-FRT-Stop -FRT-rpr (DEMISE) plasmid | this paper | progenitor: pUAST-FRT-Stop-FRT-mCD8-GFP | |
| recombinant DNA reagent | pUAST-FRT-Stop- FRT-mCD8-GFP plasmid | Potter et al., 2010 | addgene #24385 | |
| commercial assay or kit | TUNEL | Roche | #12156792910 |
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.38327.014
