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Injury-stimulated Hedgehog signaling promotes regenerative proliferation of Drosophila intestinal stem cells.

Tian A, Shi Q, Jiang A, Li S, Wang B, Jiang J - J. Cell Biol. (2015)

Bottom Line: Many adult tissues are maintained by resident stem cells that elevate their proliferation in response to injury.Here we show that injury induces Hedgehog (Hh) signaling in enteroblasts (EBs) to promote intestinal stem cell (ISC) proliferation in Drosophila melanogaster adult midgut.Hence, our study uncovers a JNK-Hh-JAK-STAT signaling axis in the regulation of regenerative stem cell proliferation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Developmental Biology and Department of Pharmacology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390.

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Ptc restricts ISC proliferation in Drosophila adult midgut. (A–K) 3- to 5-d-old adult female flies were heat shocked at 37°C for 1 h to induce MACRM clones for control FRT chromosomes (A, A′, F, F′, I, and I′), FRT42 ptcIIW (B and B′), FRT42 ptcS2 (C and C′), FRT42 cos22 (D and D′), or FRT40 smo3 chromosome (G, G′, J, and J′) and cultured at 18°C for 10 (A–H) or 20 d ACI (I–K). Midguts were immunostained for GFP (green) and DRAQ5 (blue) and clone size was quantified for each genotype (E, H, and K). Clones were marked by GFP and nuclei were marked by DRAQ5. Only ISC lineage clones in the posterior region of midguts were included for quantification. Bars, 50 µm. Data are mean ± SEM. n = 150. ***, P < 0.001. ns, not significant.
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fig1: Ptc restricts ISC proliferation in Drosophila adult midgut. (A–K) 3- to 5-d-old adult female flies were heat shocked at 37°C for 1 h to induce MACRM clones for control FRT chromosomes (A, A′, F, F′, I, and I′), FRT42 ptcIIW (B and B′), FRT42 ptcS2 (C and C′), FRT42 cos22 (D and D′), or FRT40 smo3 chromosome (G, G′, J, and J′) and cultured at 18°C for 10 (A–H) or 20 d ACI (I–K). Midguts were immunostained for GFP (green) and DRAQ5 (blue) and clone size was quantified for each genotype (E, H, and K). Clones were marked by GFP and nuclei were marked by DRAQ5. Only ISC lineage clones in the posterior region of midguts were included for quantification. Bars, 50 µm. Data are mean ± SEM. n = 150. ***, P < 0.001. ns, not significant.

Mentions: As an initial step to investigate whether Hh signaling plays a role in Drosophila adult midgut homeostasis, we generated GFP-labeled homozygous clones for ptc or smo mutations in 3- to 5-d-old adult females using the mosaic analysis with a repressible cell marker (MARCM) system (Lee and Luo, 2001). Two ptc alleles, ptcIIW (a allele) and ptcS2 (a strong allele), and smo3 (a allele) were used. After clone induction, adult flies were cultured at 18°C for certain periods of time before midguts were dissected for immunostaining. 10 d after clone induction (ACI), the majority of the control ISC lineage clones in the posterior region of midguts contained three to five cells whereas the majority of ptc mutant ISC lineage clones contained more than five cells (Fig. 1, A–C′ and E). Of note, we only included ISC lineage clones in the posterior region of midguts for quantification because of the regional difference in ISC proliferation rate (Buchon et al., 2013; Marianes and Spradling, 2013). The observed increase in clone size suggests that ptc mutant clones proliferated faster than the control clones, a notion that was confirmed by examining the mitotic index in midguts carrying ptc mutant clones or in which ptc was inactivated by RNAi (see results in Fig. 3).


Injury-stimulated Hedgehog signaling promotes regenerative proliferation of Drosophila intestinal stem cells.

Tian A, Shi Q, Jiang A, Li S, Wang B, Jiang J - J. Cell Biol. (2015)

Ptc restricts ISC proliferation in Drosophila adult midgut. (A–K) 3- to 5-d-old adult female flies were heat shocked at 37°C for 1 h to induce MACRM clones for control FRT chromosomes (A, A′, F, F′, I, and I′), FRT42 ptcIIW (B and B′), FRT42 ptcS2 (C and C′), FRT42 cos22 (D and D′), or FRT40 smo3 chromosome (G, G′, J, and J′) and cultured at 18°C for 10 (A–H) or 20 d ACI (I–K). Midguts were immunostained for GFP (green) and DRAQ5 (blue) and clone size was quantified for each genotype (E, H, and K). Clones were marked by GFP and nuclei were marked by DRAQ5. Only ISC lineage clones in the posterior region of midguts were included for quantification. Bars, 50 µm. Data are mean ± SEM. n = 150. ***, P < 0.001. ns, not significant.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4362464&req=5

fig1: Ptc restricts ISC proliferation in Drosophila adult midgut. (A–K) 3- to 5-d-old adult female flies were heat shocked at 37°C for 1 h to induce MACRM clones for control FRT chromosomes (A, A′, F, F′, I, and I′), FRT42 ptcIIW (B and B′), FRT42 ptcS2 (C and C′), FRT42 cos22 (D and D′), or FRT40 smo3 chromosome (G, G′, J, and J′) and cultured at 18°C for 10 (A–H) or 20 d ACI (I–K). Midguts were immunostained for GFP (green) and DRAQ5 (blue) and clone size was quantified for each genotype (E, H, and K). Clones were marked by GFP and nuclei were marked by DRAQ5. Only ISC lineage clones in the posterior region of midguts were included for quantification. Bars, 50 µm. Data are mean ± SEM. n = 150. ***, P < 0.001. ns, not significant.
Mentions: As an initial step to investigate whether Hh signaling plays a role in Drosophila adult midgut homeostasis, we generated GFP-labeled homozygous clones for ptc or smo mutations in 3- to 5-d-old adult females using the mosaic analysis with a repressible cell marker (MARCM) system (Lee and Luo, 2001). Two ptc alleles, ptcIIW (a allele) and ptcS2 (a strong allele), and smo3 (a allele) were used. After clone induction, adult flies were cultured at 18°C for certain periods of time before midguts were dissected for immunostaining. 10 d after clone induction (ACI), the majority of the control ISC lineage clones in the posterior region of midguts contained three to five cells whereas the majority of ptc mutant ISC lineage clones contained more than five cells (Fig. 1, A–C′ and E). Of note, we only included ISC lineage clones in the posterior region of midguts for quantification because of the regional difference in ISC proliferation rate (Buchon et al., 2013; Marianes and Spradling, 2013). The observed increase in clone size suggests that ptc mutant clones proliferated faster than the control clones, a notion that was confirmed by examining the mitotic index in midguts carrying ptc mutant clones or in which ptc was inactivated by RNAi (see results in Fig. 3).

Bottom Line: Many adult tissues are maintained by resident stem cells that elevate their proliferation in response to injury.Here we show that injury induces Hedgehog (Hh) signaling in enteroblasts (EBs) to promote intestinal stem cell (ISC) proliferation in Drosophila melanogaster adult midgut.Hence, our study uncovers a JNK-Hh-JAK-STAT signaling axis in the regulation of regenerative stem cell proliferation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Developmental Biology and Department of Pharmacology, University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390.

Show MeSH
Related in: MedlinePlus