Letter | Published:

Tricellular junctions regulate intestinal stem cell behaviour to maintain homeostasis

Nature Cell Biology volume 19, pages 5259 (2017) | Download Citation

Abstract

Ageing results in loss of tissue homeostasis across taxa1. In the intestine of Drosophila melanogaster, ageing is correlated with an increase in intestinal stem cell (ISC) proliferation, a block in terminal differentiation of progenitor cells, activation of inflammatory pathways, and increased intestinal permeability2. However, causal relationships between these phenotypes remain unclear. Here, we demonstrate that ageing results in altered localization and expression of septate junction proteins in the posterior midgut, which is quite pronounced in differentiated enterocytes (ECs) at tricellular junctions (TCJs). Acute loss of the TCJ protein Gliotactin (Gli) in ECs results in increased ISC proliferation and a block in differentiation in intestines from young flies, demonstrating that compromised TCJ function is sufficient to alter ISC behaviour in a non-autonomous manner. Blocking the Jun N-terminal kinase signalling pathway is sufficient to suppress changes in ISC behaviour, but has no effect on loss of intestinal barrier function, as a consequence of Gli depletion. Our work demonstrates a pivotal link between TCJs, stem cell behaviour, and intestinal homeostasis and provides insights into causes of age-onset and gastrointestinal diseases.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Accessions

Primary accessions

Gene Expression Omnibus

References

  1. 1.

    & Emerging models and paradigms for stem cell ageing. Nat. Cell Biol. 13, 506–512 (2011).

  2. 2.

    & Gastrointestinal stem cells in health and disease: from flies to humans. Dis. Models Mech. 9, 487–499 (2016).

  3. 3.

    , & Epithelial barriers in homeostasis and disease. Annu. Rev. Pathol. 5, 119–144 (2010).

  4. 4.

    & Evidence that stem cells reside in the adult Drosophila midgut epithelium. Nature 439, 475–479 (2006).

  5. 5.

    & The adult Drosophila posterior midgut is maintained by pluripotent stem cells. Nature 439, 470–474 (2006).

  6. 6.

    , & JNK activity in somatic stem cells causes loss of tissue homeostasis in the aging Drosophila gut. Cell Stem Cell 3, 442–455 (2008).

  7. 7.

    et al. Cytokine/Jak/Stat signaling mediates regeneration and homeostasis in the Drosophila midgut. Cell 137, 1343–1355 (2009).

  8. 8.

    et al. Modulation of longevity and tissue homeostasis by the Drosophila PGC-1 homolog. Cell Metab. 14, 623–634 (2011).

  9. 9.

    , & Intestinal barrier dysfunction links metabolic and inflammatory markers of aging to death in Drosophila. Proc. Natl Acad. Sci. USA 109, 21528–21533 (2012).

  10. 10.

    & The development of cellular junctions in the Drosophila embryo. Dev. Biol. 161, 563–596 (1994).

  11. 11.

    Septate junctions in imaginal disks of Drosophila: a model for the redistribution of septa during cell rearrangement. J. Cell Biol. 94, 77–87 (1982).

  12. 12.

    , & Gliotactin, a novel marker of tricellular junctions, is necessary for septate junction development in Drosophila. J. Cell Biol. 161, 991–1000 (2003).

  13. 13.

    et al. The triple-repeat protein anakonda controls epithelial tricellular junction formation in Drosophila. Dev. Cell 33, 535–548 (2015).

  14. 14.

    et al. Bark beetle controls epithelial morphogenesis by septate junction maturation in Drosophila. Dev. Biol. 400, 237–247 (2015).

  15. 15.

    , , & Gliotactin, a novel transmembrane protein on peripheral glia, is required to form the blood-nerve barrier in Drosophila. Cell 81, 757–767 (1995).

  16. 16.

    et al. Gliotactin and Discs large form a protein complex at the tricellular junction of polarized epithelial cells in Drosophila. J. Cell Sci. 119, 4391–4401 (2006).

  17. 17.

    , , & Gliotactin and Discs large are co-regulated to maintain epithelial integrity. J. Cell Sci. 126, 1134–1143 (2013).

  18. 18.

    , & Extending healthy life span–from yeast to humans. Science 328, 321–326 (2010).

  19. 19.

    , , & A conditional tissue-specific transgene expression system using inducible GAL4. Proc. Natl Acad. Sci. USA 98, 12596–12601 (2001).

  20. 20.

    , , & P[Switch], a system for spatial and temporal control of gene expression in Drosophila melanogaster. Proc. Natl Acad. Sci. USA 98, 12602–12607 (2001).

  21. 21.

    et al. Age-related upregulation of Drosophila caudal gene via NF-κB in the adult posterior midgut. Biochim. Biophys. Acta 1780, 1093–1100 (2008).

  22. 22.

    , & The role of p38b MAPK in age-related modulation of intestinal stem cell proliferation and differentiation in Drosophila. Aging 1, 637–651 (2009).

  23. 23.

    , , & Invasive and indigenous microbiota impact intestinal stem cell activity through multiple pathways in Drosophila. Genes Dev. 23, 2333–2344 (2009).

  24. 24.

    , & Gut homeostasis in a microbial world: insights from Drosophila melanogaster. Nat. Rev. Microbiol. 11, 615–626 (2013).

  25. 25.

    , , & PGRP-SC2 promotes gut immune homeostasis to limit commensal dysbiosis and extend lifespan. Cell 156, 109–122 (2014).

  26. 26.

    et al. Distinct shifts in microbiota composition during Drosophila aging impair intestinal function and drive mortality. Cell Rep. 12, 1656–1667 (2015).

  27. 27.

    , , & Regulation of Drosophila lifespan by JNK signaling. Exp. Gerontol. 46, 349–354 (2011).

  28. 28.

    & Warts and Yorkie mediate intestinal regeneration by influencing stem cell proliferation. Curr. Biol. 20, 1580–1587 (2010).

  29. 29.

    & The intestinal epithelial barrier: does it become impaired with age? Dig. Dis. 27, 240–245 (2009).

  30. 30.

    et al. Age-related changes in small intestinal mucosa epithelium architecture and epithelial tight junction in rat models. Aging Clin. Exp. Res. 26, 183–191 (2014).

  31. 31.

    & Age-associated remodeling of the intestinal epithelial barrier. J. Gerontol. A Biol. Sci. Med. Sci. 68, 1045–1056 (2013).

  32. 32.

    , & Spatiotemporal gene expression targeting with the TARGET and gene-switch systems in Drosophila. Sci. STKE 2004, pl6 (2004).

  33. 33.

    et al. Puckered encodes a phosphatase that mediates a feedback loop regulating JNK activity during dorsal closure in Drosophila. Genes Dev. 12, 557–570 (1998).

  34. 34.

    , , & A gain-of-function screen identifying genes required for growth and pattern formation of the Drosophila melanogaster wing. Genetics 183, 1005–1026 (2009).

  35. 35.

    et al. Features of endogenous cardiomyocyte chromatin revealed by super-resolution STED microscopy. J. Mol. Cell. Cardiol. 53, 552–558 (2012).

  36. 36.

    CellProfiler: image analysis software for identifying and quantifying cell phenotypes. Genome Biol. 7, R100 (2006).

  37. 37.

    The persistence of a microbial flora during postembryogenesis of Drosophila melanogaster. J. Invertebr. Pathol. 14, 365–374 (1969).

Download references

Acknowledgements

The authors thank H. Jasper (The Buck Institute for Research on Aging, USA), V. Auld (University of British Columbia, Canada), J. F. de Celis (CBMSO, Spain), M. Furuse (Kobe University, Japan), the Vienna Drosophila RNAi Center (VDRC), and Bloomington Stock Center for reagents, M. Cilluffo from the Brain Research Institute at UCLA EM core facility, and the Jones and Walker laboratories for comments on the manuscript. In addition, we would like to thank Y. Wu and E. Stefani for their sharing their time, expertise, and STED technology. This work was supported by the Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research at the University of California, Los Angeles and the Rose Hills Foundation (D.L.J.), and the NIH: AG028092 (D.L.J.), AG040288 (D.L.J. and D.W.W.), AG049157 (D.W.W.), and a training grant that supports the UCLA-Caltech Medical Scientist Training Program, GM08042 (S.L.).

Author information

Author notes

    • Rebecca I. Clark

    Present address: School of Biological and Biomedical Sciences, Durham University, Durham DH1 3LE, UK.

Affiliations

  1. Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, California 90095, USA

    • Martin Resnik-Docampo
    • , Christopher L. Koehler
    • , Vivien Sauer
    • , Daniel M. Wong
    • , Sophia Lewis
    • , Cecilia D’Alterio
    •  & D. Leanne Jones
  2. Department of Integrative Biology and Physiology, University of California, Los Angeles, California 90095, USA

    • Rebecca I. Clark
    • , Joseph M. Schinaman
    •  & David W. Walker
  3. Molecular Biology Institute, University of California, Los Angeles, California 90095, USA

    • David W. Walker
    •  & D. Leanne Jones
  4. Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, California 90095, USA

    • D. Leanne Jones

Authors

  1. Search for Martin Resnik-Docampo in:

  2. Search for Christopher L. Koehler in:

  3. Search for Rebecca I. Clark in:

  4. Search for Joseph M. Schinaman in:

  5. Search for Vivien Sauer in:

  6. Search for Daniel M. Wong in:

  7. Search for Sophia Lewis in:

  8. Search for Cecilia D’Alterio in:

  9. Search for David W. Walker in:

  10. Search for D. Leanne Jones in:

Contributions

M.R.-D. designed, performed and analysed experiments and wrote the manuscript. C.L.K., R.I.C., J.M.S., D.M.W., V.S., S.L. and C.D’A. designed, performed and analysed experiments. D.W.W. designed and analysed experiments. D.L.J. designed and analysed experiments and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to D. Leanne Jones.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Information

Excel files

  1. 1.

    Supplementary Table 1

    Supplementary Information

  2. 2.

    Supplementary Table 2

    Supplementary Information

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ncb3454

Further reading