Letter | Published:

Tissue damage detection by osmotic surveillance

Nature Cell Biology volume 15, pages 11231130 (2013) | Download Citation

Abstract

How tissue damage is detected to induce inflammatory responses is unclear. Most studies have focused on damage signals released by cell breakage and necrosis1. Whether tissues use other cues in addition to cell lysis to detect that they are damaged is unknown. We find that osmolarity differences between interstitial fluid and the external environment mediate rapid leukocyte recruitment to sites of tissue damage in zebrafish by activating cytosolic phospholipase a2 (cPLA2) at injury sites. cPLA2 initiates the production of non-canonical arachidonate metabolites that mediate leukocyte chemotaxis through a 5-oxo-ETE receptor (OXE-R). Thus, tissues can detect damage through direct surveillance of barrier integrity, with cell swelling probably functioning as a pro-inflammatory intermediate in the process.

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References

  1. 1.

    & Hydrophobicity: an ancient damage-associated molecular pattern that initiates innate immune responses. Nat. Rev. Immunol. 4, 469–478 (2004).

  2. 2.

    et al. Electrical signals control wound healing through phosphatidylinositol-3-OH kinase-gamma and PTEN. Nature 442, 457–460 (2006).

  3. 3.

    , & Increased bronchial chloride concentration in cystic fibrosis. Scand. J. Clin. Lab Invest. 49, 121–124 (1989).

  4. 4.

    , & Elemental composition of human airway surface fluid in healthy and diseased airways. Am. Rev. Respir. Dis. 148, 1633–1637 (1993).

  5. 5.

    , , , & Wound healing and inflammation: embryos reveal the way to perfect repair. Phil. Trans. R. Soc. Lond. Ser. B 359, 777–784 (2004).

  6. 6.

    & Reverse leukocyte migration can be attractive or repulsive. Trends Cell Biol. 18, 298–306 (2008).

  7. 7.

    & A model 450 million years in the making: zebrafish and vertebrate immunity. Dis. Models Mech. 5, 38–47 (2012).

  8. 8.

    & Fish immunology. Curr. Biol. 19, R678–R682 (2009).

  9. 9.

    et al. in Molecular Biology of the Cell 4th edn (ed. Redd, M. J.) (Garland Science, 2002).

  10. 10.

    et al. Resolution of inflammation by retrograde chemotaxis of neutrophils in transgenic zebrafish. J. Leukoc. Biol. 80, 1281–1288 (2006).

  11. 11.

    The role of osmotic pressure in the analogy between solutions and gases. J. Membr. Sci. 100, 39–44 (1995).

  12. 12.

    , & Physiology of cell volume regulation in vertebrates. Phys. Rev. 89, 193–277 (2009).

  13. 13.

    , , & A tissue-scale gradient of hydrogen peroxide mediates rapid wound detection in zebrafish. Nature 459, 996–999 (2009).

  14. 14.

    , , & Lyn is a redox sensor that mediates leukocyte wound attraction in vivo. Nature 480, 109–112 (2011).

  15. 15.

    , , , & Prioritization of competing damage and developmental signals by migrating macrophages in the Drosophila embryo. Curr. Biol. 20, 464–470 (2010).

  16. 16.

    , , , & Live imaging of innate immune cell sensing of transformed cells in zebrafish larvae: parallels between tumor initiation and wound inflammation. PLoS Biol. 8, e1000562 (2010).

  17. 17.

    & Phospholipase A2 structure/function, mechanism, and signaling. J. Lipid Res. 50 (suppl.), S237–S242 (2009).

  18. 18.

    et al. Translocation of the 85-kDa phospholipase A2 from cytosol to the nuclear envelope in rat basophilic leukemia cells stimulated with calcium ionophore or IgE/antigen. J. Biol. Chem. 270, 15359–15367 (1995).

  19. 19.

    , , & Translocation of cytosolic phospholipase A2 to the nuclear envelope elicits topographically localized phospholipid hydrolysis. Biochem. J. 318, 797–803 (1996).

  20. 20.

    , & ‘Cytosolic’ phospholipase A2 is in the nucleus of subconfluent endothelial cells but confined to the cytoplasm of confluent endothelial cells and redistributes to the nuclear envelope and cell junctions upon histamine stimulation. Lab. Invest. 74, 684–695 (1996).

  21. 21.

    , , & Nuclear localisation of cytosolic phospholipase A2-alpha in the EA.hy.926 human endothelial cell line is proliferation dependent and modulated by phosphorylation. J. Cell Sci. 115, 4533–4543 (2002).

  22. 22.

    et al. Coordinate development of skin cells and cutaneous sensory axons in zebrafish. J. Comp. Neurol. 520, 816–831 (2012).

  23. 23.

    et al. In vivo cell and tissue dynamics underlying zebrafish fin fold regeneration. PLoS One 7, e51766 (2012).

  24. 24.

    , , , & Cell swelling activates phospholipase A2 in Ehrlich ascites tumor cells. J. Membr. Biol. 160, 47–58 (1997).

  25. 25.

    , , , & Swelling-induced arachidonic acid release via the 85-kDa cPLA2 in human neuroblastoma cells. J. Neurophysiol. 79, 1441–1449 (1998).

  26. 26.

    , , & Hypotonic cell swelling induces translocation of the alpha isoform of cytosolic phospholipase A2 but not the γ isoform in Ehrlich ascites tumor cells. Eur. J. Biochem. 267, 5531–5539 (2000).

  27. 27.

    et al. Cell swelling, heat, and chemical agonists use distinct pathways for the activation of the cation channel TRPV4. Proc. Natl Acad. Sci. USA 101, 396–401 (2004).

  28. 28.

    , , & A mechanosensitive ion channel regulating cell volume. Am. J. Physiol. Cell Physiol. 298, C1424–C1430 (2010).

  29. 29.

    , , & Calcium flashes orchestrate the wound inflammatory response through DUOX activation and hydrogen peroxide release. Curr. Biol. 23, 424–429 (2013).

  30. 30.

    , , , & Long-range Ca2+ waves transmit brain-damage signals to microglia. Dev. Cell 22, 1138–1148 (2012).

  31. 31.

    et al. Evidence of 5-lipoxygenase overexpression in the skin of patients with systemic sclerosis: a newly identified pathway to skin inflammation in systemic sclerosis. Arthritis Rheum. 44, 1865–1875 (2001).

  32. 32.

    , & Leukotriene synthesis by epithelial cells. Hist. Histopathol. 18, 587–595 (2003).

  33. 33.

    et al. Global analysis of the haematopoietic and endothelial transcriptome during zebrafish development. Mech. Dev. 130, 122–131 (2013).

  34. 34.

    , & 5-Oxo-ETE and the OXE receptor. Prostaglandins 89, 98–104 (2009).

  35. 35.

    , & 5-Oxo-6,8,11,14-eicosatetraenoic acid is a potent stimulator of human eosinophil migration. J. Immunol. 154, 4123–4132 (1995).

  36. 36.

    , & Hydrogen peroxide-induced chemotaxis of mouse peritoneal neutrophils. Eur. J. Cell Biol. 70, 347–351 (1996).

  37. 37.

    , , & Rac regulates PtdInsP(3) signaling and the chemotactic compass through a redox-mediated feedback loop. Blood 118, 6164–6171 (2011).

  38. 38.

    et al. Impaired cell volume regulation in intestinal crypt epithelia of cystic fibrosis mice. Proc. Natl Acad. Sci. USA 92, 9038–9041 (1995).

  39. 39.

    & The cell biology of acute myocardial ischemia. Annu. Rev. Med. 42, 225–246 (1991).

  40. 40.

    et al. Evidence for periciliary liquid layer depletion, not abnormal ion composition, in the pathogenesis of cystic fibrosis airways disease. Cell 95, 1005–1015 (1998).

  41. 41.

    et al. Transparent adult zebrafish as a tool for in vivo transplantation analysis. Cell Stem Cell 2, 183–189 (2008).

  42. 42.

    & Zebrafish: A Practical Approach (Oxford Univ. Press, 2002).

  43. 43.

    et al. An expanded palette of genetically encoded Ca(2)(+) indicators. Science 333, 1888–1891 (2011).

  44. 44.

    et al. The Tol2kit: a multisite gateway-based construction kit for Tol2 transposon transgenesis constructs. Dev. Dyn. 236, 3088–3099 (2007).

  45. 45.

    , , , & The zebrafish lysozyme C promoter drives myeloid-specific expression in transgenic fish. BMC Dev. Biol. 7, 42 (2007).

  46. 46.

    , & A zebrafish histone variant H2A.F/Z and a transgenic H2A.F/Z:GFP fusion protein for in vivo studies of embryonic development. Dev. Genes Evol. 211, 603–610 (2001).

  47. 47.

    et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).

  48. 48.

    et al. The lta4h locus modulates susceptibility to mycobacterial infection in zebrafish and humans. Cell 140, 717–730 (2010).

  49. 49.

    et al. p53 activation by knockdown technologies. PLoS Gen. 3, e78 (2007).

  50. 50.

    et al. Definitive hematopoiesis initiates through a committed erythromyeloid progenitor in the zebrafish embryo. Development 134, 4147–4156 (2007).

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Acknowledgements

We thank T. Mitchison, A. Hall, M. Overholtzer and A. Kapus for their valuable thoughts and suggestions on the manuscript. This work was supported by NIH grant GM099970 and an L. V. Gerstner Young Investigator award.

Author information

Affiliations

  1. Cell Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA

    • Balázs Enyedi
    • , Snigdha Kala
    • , Tijana Nikolich-Zugich
    •  & Philipp Niethammer

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Contributions

P.N. conceived the project. B.E. and P.N. designed the experiments. B.E., P.N., S.K. and T.N-Z. carried out the experiments. P.N. and B.E. wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Philipp Niethammer.

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  1. 1.

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    Supplementary Information

Videos

  1. 1.

    Hypotonicity is required for rapid leukocyte recruitment to larval zebrafish tail fin wounds.

    This movie shows leukocyte recruitment after tail fin wounding of wt larvae in hypotonic (control) or isotonic (145 mM NaCl) medium. Imaging starts 3 min pw (60 s per frame). Scale bar, 100 μm.

  2. 2.

    Isotonicity reversibly inhibits rapid leukocyte recruitment to larval zebrafish tail fin wounds.

    This movie shows leukocyte recruitment after tail fin wounding of wt larvae in isotonic medium that was shifted to hypotonic medium (‘i → h’ shift) or isotonic medium (‘i → i’ shift). Imaging starts 3 min pw (30 sec per frame). Scale bar, 100 μm.

  3. 3.

    Hypotonicity locally activates cPLA2 at the wound site.

    This movie montage shows cPLA2–mKate2 translocation to the nuclear membrane induced by UV-laser wounding in hypotonic (hypo), isotonic (iso) or hypotonic medium supplemented with 500 μM Gd3+(hypo+Gd3+). Laser wounding at 1 min (15 sec per frame). Scale bar, 100 μm.

  4. 4.

    Gd3+ exposure enhances wound margin swelling.

    This movie shows leukocyte recruitment after hypotonic tail fin wounding of WT larvae in the presence or absence of 500 μM Gd3+. Imaging starts 3 min pw, Gd3+ added at 0 min (60 sec per frame). Scale bar, 100 μm.

  5. 5.

    Extracellular Ca2+ is required for cPLA2 activation.

    This movie montage shows cPLA2–mKate2 translocation to the nuclear membrane induced by calcium-switch in hypotonic medium. Hypotonic Ca2+-free medium supplemented with 1mM EGTA was added at 0 min to the larvae that were prewounded in isotonic medium with EGTA. Larve were imaged for 40 min with (A, C) or without (B) re-addition of Ca2+ at 5 min. Movies from different experiments. Low magnification scale bar, 100 μm; high magnification scale bar, 10 μm (15 sec per frame).

  6. 6.

    Tail fin injury rapidly increases cytoplasmic [Ca2+ at the wound site irrespective of medium tonicity.

    This movie shows the cytosolic Ca2+ signal induced by UV-laser wounding of larval zebrafish tail fins maintained in hypotonic (h) or isotonic (i) medium. Laser wounding at 19 sec (3 sec per frame). Scale bar, 100 μm.

  7. 7.

    cPLA2 is required for rapid leukocyte recruitment to larval zebrafish tail fin wounds.

    This movie shows leukocyte recruitment in cpla2 morphant larva (cpla2 MO) versus WT larva 3dpf. Imaging starts 3 min pw (60 sec per frame). Scale bar, 100 μm.

  8. 8.

    5-KETE is a chemoattractant for leukocytes in zebrafish.

    This movie shows leukocyte recruitment after isotonic tail fin wounding of WT larvae in the presence or absence of 2 μM 5-KETE (5-oxo-ETE). Imaging starts 3 min pw, 5-KETE added at 0 min (60 sec per frame). Scale bar, 100 μm.

  9. 9.

    OXE-R is required for rapid leukocyte recruitment to larval zebrafish tail fin wounds.

    This movie shows leukocyte recruitment in oxer1 morphant larva (oxer1 MO) versus WT larva 3dpf. Imaging starts 3 min pw (60 sec per frame). Scale bar, 100 μm.

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DOI

https://doi.org/10.1038/ncb2818

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