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Electrical signals control wound healing through phosphatidylinositol-3-OH kinase-γ and PTEN

Abstract

Wound healing is essential for maintaining the integrity of multicellular organisms. In every species studied, disruption of an epithelial layer instantaneously generates endogenous electric fields, which have been proposed to be important in wound healing1,2,3. The identity of signalling pathways that guide both cell migration to electric cues and electric-field-induced wound healing have not been elucidated at a genetic level. Here we show that electric fields, of a strength equal to those detected endogenously, direct cell migration during wound healing as a prime directional cue. Manipulation of endogenous wound electric fields affects wound healing in vivo. Electric stimulation triggers activation of Src and inositol–phospholipid signalling, which polarizes in the direction of cell migration. Notably, genetic disruption of phosphatidylinositol-3-OH kinase-γ (PI(3)Kγ) decreases electric-field-induced signalling and abolishes directed movements of healing epithelium in response to electric signals. Deletion of the tumour suppressor phosphatase and tensin homolog (PTEN) enhances signalling and electrotactic responses. These data identify genes essential for electrical-signal-induced wound healing and show that PI(3)Kγ and PTEN control electrotaxis.

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Figure 1: Electrical signals direct cell migration in wound healing and activate selected signalling pathways.
Figure 2: Electrotaxis requires PI(3)Kγ.
Figure 3: The tumour suppressor PTEN negatively regulates electrotaxis.
Figure 4: PI(3)Kγ is required for electrotactic cell movement in wound healing of stratified epithelium in ex vivo cornea cultures.

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Acknowledgements

This work was supported by grants from the Wellcome Trust and Royal Society (to M.Z.), the Institute of Molecular Biotechnology of Austria of the Austrian Academy of Sciences, the Austria Ministry of Sciences, the Austrian National Bank, and a European Union Framework 6 excellence grant (all to J.M.P.). We thank all members of the Zhao, Bourne and Penninger laboratories for discussions and technical assistance. Author Contributions M.Z. designed the experiments, took part in the cell migration and western blotting experiments, analysed the results and wrote the paper. J.M.P. designed the genetic analysis of the electric-field-induced signalling pathway, analysed the data and wrote the paper. B.S. did the in vivo experiments, most experiments with cells and tissues from transgenic mice. B.S. and J.P. did most of the cell migration and wound healing assays. T.W. performed the first signalling experiment and genotyping. B.R. performed the vibrating probe measurements. G.T., F.W. and P.W. did the experiments with HL60 cells. B.S., A.G. and Y.G. did the experiments on fibroblasts. P.N.D., A.S. and T.S. provided mouse and D. discoideum lines essential for the experiments. J.V.F., H.B. and C.D.M. helped with some of the experimental design, writing and analysis of the data. All authors discussed the results and commented on the manuscript.

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Correspondence to Min Zhao or Josef M. Penninger.

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Supplementary information

Supplementary Notes

This file contains the Supplementary Introduction, Supplementary Methods, Supplementary Results, Supplementary Figures 1-17, Supplementary Movie Legends and additional references (PDF 1387 kb)

Supplementary Movie 1

The electric signal is a predominant cue directing cell migration in monolayer wound healing assays. (MOV 9353 kb)

Supplementary Movie 2

The electrical signal directs migration of primary neutrophils and HL60 cells. (MOV 7578 kb)

Supplementary Movie 3

Impaired electrotactic responses of p110γ-/- fibroblasts. (MOV 8971 kb)

Supplementary Movie 4

The electric signal is a predominant cue directing cell migration in stratified epithelial wound healing. (MOV 9912 kb)

Supplementary Movie 5

Polarized PI3K signaling during neutrophil electrotaxis. (MOV 3617 kb)

Supplementary Movie 6

Chemotactic sensing is not required for electrotactic responses in Dictyostelium. (MOV 3356 kb)

Supplementary Movie 7

Electrotaxis of keratinocytes requires PI3Kγ. (MOV 6989 kb)

Supplementary Movie 8

PI3Kγ is required for electrotactic cell migration in monolayer wound healing assays. (MOV 9315 kb)

Supplementary Movie 9

Impaired electrotactic responses of p110γ-/- neutrophils. (MOV 6201 kb)

Supplementary Movie 10

Wortmannin significantly inhibits electrotactic migration of mouse keratinocytes. (MOV 9683 kb)

Supplementary Movie 11

Wortmannin significantly inhibits electrotactic migration of keratinocytes in monolayer wound healing assays. (MOV 9514 kb)

Supplementary Movie 12

The tumor suppressor PTEN is a negative regulator of the electrotactic response. (MOV 5793 kb)

Supplementary Movie 13

Loss of pten gene significantly enhances the electrotactic response of keratinocyte in monolayer wound healing assays. (MOV 9551 kb)

Supplementary Movie 14

PI3Kγ is essential for electrotactic migration of stratified epithelial cells in wound healing. (MOV 9892 kb)

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Zhao, M., Song, B., Pu, J. et al. Electrical signals control wound healing through phosphatidylinositol-3-OH kinase-γ and PTEN. Nature 442, 457–460 (2006). https://doi.org/10.1038/nature04925

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