Letter | Published:

Lyn is a redox sensor that mediates leukocyte wound attraction in vivo

Nature volume 480, pages 109112 (01 December 2011) | Download Citation

Abstract

Tissue wounding induces the rapid recruitment of leukocytes1. Wounds and tumours—a type of ‘unhealed wound’2—generate hydrogen peroxide (H2O2) through an NADPH oxidase (NOX). This extracellular H2O2 mediates recruitment of leukocytes, particularly the first responders of innate immunity, neutrophils, to injured tissue3,4,5,6. However, the sensor that neutrophils use to detect the redox state at wounds is unknown. Here we identify the Src family kinase (SFK) Lyn as a redox sensor that mediates initial neutrophil recruitment to wounds in zebrafish larvae. Lyn activation in neutrophils is dependent on wound-derived H2O2 after tissue injury, and inhibition of Lyn attenuates neutrophil wound recruitment. Inhibition of SFKs also disrupted H2O2-mediated chemotaxis of primary human neutrophils. In vitro analysis identified a single cysteine residue, C466, as being responsible for direct oxidation-mediated activation of Lyn. Furthermore, transgenic-tissue-specific reconstitution with wild-type Lyn and a cysteine mutant revealed that Lyn C466 is important for the neutrophil wound response and downstream signalling in vivo. This is the first identification, to our knowledge, of a physiological redox sensor that mediates leukocyte wound attraction in multicellular organisms.

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Acknowledgements

We thank K. T. Chan for help with tissue culture work, J. M. Green and P.-Y. Lam for help with in situ hybridization, M. Shelef and S. Wernimont for drawing blood, and A. J. Wiemer for insightful discussion and critical reading of the manuscript. This work was supported by American Heart Association fellowship 11PRE4890041 (S.K.Y.), National Institutes of Health Grant GM074827 (A.H.), NIH Research Training Grant in Hematology 5T32 HL07899 (T.W.S.) and UW MSTP (T.W.S.).

Author information

Author notes

    • Taylor W. Starnes
    •  & Qing Deng

    These authors contributed equally to this work.

Affiliations

  1. Program in Cellular and Molecular Biology, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA

    • Sa Kan Yoo
  2. Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA

    • Taylor W. Starnes
    • , Qing Deng
    •  & Anna Huttenlocher
  3. Department of Pediatrics, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA

    • Anna Huttenlocher

Authors

  1. Search for Sa Kan Yoo in:

  2. Search for Taylor W. Starnes in:

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Contributions

S.K.Y. designed the research, performed the experiments, analysed data and wrote the paper. T.W.S. contributed to development and data analysis of the in vitro chemotaxis assay. Q.D. constructed the HyPer probe and contributed expertise in zebrafish injection. A.H. designed the research, analysed data and co-wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Anna Huttenlocher.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    This file contains Supplementary Figures 1-13 with legends, legends for Supplementary Movies 1-4 and additional references.

Videos

  1. 1.

    Supplementary Movie 1

    This movie shows time-lapse imaging of H2O2 immediately after tail transection in 2.5 dpf larvae expressing HyPer probe - see Supplementary Information file for full legend.

  2. 2.

    Supplementary Movie 2

    This movie shows time-lapse imaging of neutrophil random migration in the cephalic mesenchyme of 3 dpf Tg(mpx:Dendra2) - see Supplementary Information file for full legend.

  3. 3.

    Supplementary Movie 3

    This movie shows four examples of representative time-lapse imaging of 2.5 dpf Tg(mpx:Dendra2) injected with lyn MO or buffer - see Supplementary Information file for full legend.

  4. 4.

    Supplementary Movie 4

    This movie shows LTB4-mediated neutrophil dissemination into the fins of zebrafish larvae - see Supplementary Information file for full legend.

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DOI

https://doi.org/10.1038/nature10632

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