Letter

Mechanical regulation of stem-cell differentiation by the stretch-activated Piezo channel

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Accepted:
Published online:

Abstract

Somatic stem cells constantly adjust their self-renewal and lineage commitment by integrating various environmental cues to maintain tissue homeostasis. Although numerous chemical and biological signals have been identified that regulate stem-cell behaviour, whether stem cells can directly sense mechanical signals in vivo remains unclear1. Here we show that mechanical stress regulates stem-cell differentiation in the adult Drosophila midgut through the stretch-activated ion channel Piezo. We find that Piezo is specifically expressed in previously unidentified enteroendocrine precursor cells, which have reduced proliferation ability and are destined to become enteroendocrine cells. Loss of Piezo activity reduces the generation of enteroendocrine cells in the adult midgut. In addition, ectopic expression of Piezo in all stem cells triggers both cell proliferation and enteroendocrine cell differentiation. Both the Piezo mutant and overexpression phenotypes can be rescued by manipulation of cytosolic Ca2+ levels, and increases in cytosolic Ca2+ resemble the Piezo overexpression phenotype, suggesting that Piezo functions through Ca2+ signalling. Further studies suggest that Ca2+ signalling promotes stem-cell proliferation and differentiation through separate pathways. Finally, Piezo is required for both mechanical activation of stem cells in a gut expansion assay and the increase of cytosolic Ca2+ in response to direct mechanical stimulus in a gut compression assay. Thus, our study demonstrates the existence of a specific group of stem cells in the fly midgut that can directly sense mechanical signals through Piezo.

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Acknowledgements

We thank R. Binari, W. Song and C. Villalta for technical support; C. Xu, S. Mohr and D. Doupe for comments on the manuscript; and G. Hasan for sharing reagents. This work was supported by the Damon Runyon Cancer Research Foundation (L.H.) and a grant from the National Institutes of Health (R21DA039582). N.P. is an investigator of the Howard Hughes Medical Institute. G.S. and A.D.T.S. are supported by the National Institutes of Health (P01GM103770).

Author information

Affiliations

  1. Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA

    • Li He
    •  & Norbert Perrimon
  2. Department of Physics, Center for Brain Science, Harvard University, Cambridge, Massachusetts 02142, USA

    • Guangwei Si
    •  & Aravinthan D. T. Samuel
  3. School of Life Science and Technology, Tongji University, Shanghai 200092, China

    • Jiuhong Huang
  4. Howard Hughes Medical Institute, Boston, Massachusetts 02115, USA

    • Norbert Perrimon

Authors

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  2. Search for Guangwei Si in:

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Contributions

J.H. and L.H. performed the initial GAL4 expression screen in fly gut. L.H. and N.P. designed the experiments. L.H. performed the Piezo-related experiments and analysed the data. G.S. and A.D.T.S. designed and fabricated the microfluidic chip and together with L.H. optimized the experimental conditions. L.H. and N.P. wrote the manuscript with input from all of the authors.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Li He or Norbert Perrimon.

Reviewer Information Nature thanks L. O’Brien, K. Venkatachalam and the other anonymous reviewer(s) for their contribution to the peer review of this work.

Publisher's note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Supplementary information

PDF files

  1. 1.

    Life Sciences Reporting Summary

Excel files

  1. 1.

    Supplementary Table 1

    This file contains a list of complete genotypes for the figures.

  2. 2.

    Supplementary Table 2

    This file contains a list of primers used for TR-qPCR.

Videos

  1. 1.

    Cytosolic Ca2+ activities in control fly midguts

    Guts from flies incubated for 4-5 days at 32 °C were dissected and imaged in live imaging medium. Z-stack images were acquired with 20 sec interval. A maximal intensity z-projection is shown in the video. Genotype of the sample: esg-Gal4, tub-Gal80ts, UAS-GFP; UAS-IVS-NES-jRGECO.

  2. 2.

    Cytosolic Ca2+ activities in Piezo over-expressing fly midguts

    Guts from flies incubated for 4-5 days at 32 °C were dissected and imaged in live imaging medium. Z-stack images were acquired with 20 sec interval. A maximal intensity z-projection is shown in the video. Genotype of the sample: esg-Gal4, tub-Gal80ts, UAS-GFP; UAS-GFP-Piezo/UAS-IVS-NES-jRGECO.

  3. 3.

    Cytosolic Ca2+ activities in midguts of flies treated with a Serca inhibitor

    Adult flies were kept on control food (5% sucrose), Thap food (5% sucrose + 0.5µM Thapsigargin), or Thap+Tram food (5% sucrose + 0.5µM Thapsigargin + 10µM Trametinib) for 4 days before imaging. Guts from flies with the indicated genotypes were dissected in live imaging medium. Z-stack images were acquired with 20 sec interval. A maximal intensity z-projection is shown in the video. Genotype of the sample: UAS- IVS-GCaMP6s / UAS-tdTomato, Dl-Gal4.

  4. 4.

    Cytosolic Ca2+ activities in midguts of flies feed on MC food

    Adult flies with indicated genotypes were fed on control (5% sucrose) or MC (5% sucrose + 10% Methylcellulose) food for 4-5 days. Midguts were dissected in live imaging medium. Z-stack images were acquired with 20 sec interval. A maximal intensity z-projection is shown in the video. Genotypes of the samples: Control, MC Fed (normal), MC Fed: UAS- IVS-GCaMP6s / UAS-tdTomato; Dl-Gal4. MC Fed, Piezo-i: UAS- IVS-GCaMP6s / UAS-tdTomato; Dl-Gal4/UAS-PiezoRNAi. MC Fed, Stim-i, IP3R-i: tubGal80ts/+; UAS- IVS-GCaMP6s / UAS-InsP3RRNAi; UAS-tdTomato, Dl-Gal4/ UAS- StimRNAi.

  5. 5.

    Cytosolic Ca2+ activities in WT and mutant of flies

    Guts from flies incubated for 5-7 days at 32 °C were dissected and imaged in live imaging medium. Z-stack images were acquired with 20 sec interval. A maximal intensity z-projection is shown in the video. Genotypes of the samples: UAS- IVS-GCaMP6s / UAS-tdTomato; Dl-Gal4. UAS- IVS-GCaMP6s / UAS-tdTomato; Dl-Gal4/UAS-PiezoRNAi. UAS- IVS-GCaMP6s, PiezoKO / PiezoKO; UAS-tdTomato /Dl-Gal4. tubGal80ts/+; UAS- IVS-GCaMP6s / UAS-tdTomato; Dl-Gal4/UAS-SercaRNAi. tubGal80ts/+; UAS- IVS-GCaMP6s / UAS- StimRNAi; UAS-tdTomato, Dl-Gal4/UAS-InsP3RRNAi.

  6. 6.

    Cytosolic Ca2+ activities in control fly midguts under mechanical compression.

    Guts from flies incubated for 5-7 days at 32 °C were dissected and imaged in live imaging medium. For each cycle, the gut was compressed for ~40 sec. Z-stack images were acquired with 2 sec interval. A maximal intensity z-projection is shown in the video. Genotype of the sample: UAS- IVS-GCaMP6s / UAS-tdTomato, Dl-Gal4.

  7. 7.

    Cytosolic Ca2+ activities in PiezoRNAi fly midguts under mechanical compression.

    Guts from flies incubated for 5-7 days at 32 °C were dissected and imaged in live imaging medium. For each cycle, the gut was compressed for ~40 sec. Z-stack images were acquired with 2 sec interval. A maximal intensity z-projection is shown in the video. Genotype of the sample: UAS- IVS-GCaMP6s / UAS-tdTomato, Dl-Gal4/ UAS-PiezoRNAi.

  8. 8.

    Cytosolic Ca2+ activities in PiezoKO fly midguts under mechanical compression.

    Guts from flies incubated for 5-7 days at 32 °C were dissected and imaged in live imaging medium. For each cycle, the gut was compressed for ~40 sec. Z-stack images were acquired with 2 sec interval. A maximal intensity z-projection is shown in the video. Genotype of the sample: UAS- IVS-GCaMP6s, PiezoKO / PiezoKO; UAS-tdTomato /Dl-Gal4.

  9. 9.

    Cytosolic Ca2+ activities in SercaRNAi fly midguts under mechanical compression

    Guts from flies incubated for 5-7 days at 32 °C were dissected and imaged in live imaging medium. For each cycle, the gut was compressed for ~40 sec. Z-stack images were acquired with 2 sec interval. A maximal intensity z-projection is shown in the video. Genotype of the sample: tubGal80ts/+; UAS- IVS-GCaMP6s / UAS-tdTomato; Dl-Gal4/UAS-SercaRNAi.

  10. 10.

    Cytosolic Ca2+ activities in StimRNAi + InsP3RRNAi fly midguts under mechanical compression

    Guts from flies incubated for 5-7 days at 32 °C were dissected and imaged in live imaging medium. For each cycle, the gut was compressed for ~40 sec. Z-stack images were acquired with 2 sec interval. A maximal intensity z-projection is shown in the video. Genotype of the sample: tubGal80ts/+; UAS- IVS-GCaMP6s / UAS- StimRNAi; UAS-tdTomato, Dl-Gal4/UAS-InsP3RRNAi.

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