Spatiotemporal dynamics of PIEZO1 localization controls keratinocyte migration during wound healing
- University of California, Irvine, United States
- The Scripps Research Institute, United States
Abstract
Keratinocytes, the predominant cell type of the epidermis, migrate to reinstate the epithelial barrier during wound healing. Mechanical cues are known to regulate keratinocyte re-epithelialization and wound healing however, the underlying molecular transducers and biophysical mechanisms remain elusive. Here, we show through molecular, cellular and organismal studies that the mechanically-activated ion channel PIEZO1 regulates keratinocyte migration and wound healing. Epidermal-specific Piezo1 knockout mice exhibited faster wound closure while gain-of-function mice displayed slower wound closure compared to littermate controls. By imaging the spatiotemporal localization dynamics of endogenous PIEZO1 channels we find that channel enrichment at some regions of the wound edge induces a localized cellular retraction that slows keratinocyte collective migration. In migrating single keratinocytes, PIEZO1 is enriched at the rear of the cell, where maximal retraction occurs, and we find that chemical activation of PIEZO1 enhances retraction during single as well as collective migration. Our findings uncover novel molecular mechanisms underlying single and collective keratinocyte migration that may suggest a potential pharmacological target for wound treatment. More broadly, we show that nanoscale spatiotemporal dynamics of Piezo1 channels can control tissue-scale events, a finding with implications beyond wound healing to processes as diverse as development, homeostasis, disease and repair.
Data availability
The datasets for graphs included in each figure have been made available as source data files.
Article and author information
Author details
Funding
National Institutes of Health (DP2AT010376)
- Medha M Pathak
National Institutes of Health (R01NS109810)
- Medha M Pathak
National Institutes of Health (R01HL143297)
- Ardem Patapoutian
Howard Hughes Medical Institute (GT11549)
- Jesse R Holt
- Medha M Pathak
George Hewitt Foundation for Medical Research
- Wei-Zheng Zeng
National Science Foundation (DMS1763272)
- Jesse R Holt
- Wei-Zheng Zeng
Simons Foundation (594598)
- Jesse R Holt
- Wei-Zheng Zeng
Howard Hughes Medical Institute
- Ardem Patapoutian
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: All studies were approved by the Institutional Animal Care and Use Committee of Uni-versity of California at Irvine (protocol number AUP-19-184) and The Scripps Research Institute (protocol number 08-0136-4), as appropriate, and performed in accordance with their guidelines.
Copyright
© 2021, Holt et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Spatiotemporal dynamics of PIEZO1 localization controls keratinocyte migration during wound healing
eLife 10:e65415.
https://doi.org/10.7554/eLife.65415
Further reading
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- Cell Biology
- Developmental Biology
Eukaryotic cells depend on exocytosis to direct intracellularly synthesized material toward the extracellular space or the plasma membrane, so exocytosis constitutes a basic function for cellular homeostasis and communication between cells. The secretory pathway includes biogenesis of secretory granules (SGs), their maturation and fusion with the plasma membrane (exocytosis), resulting in release of SG content to the extracellular space. The larval salivary gland of Drosophila melanogaster is an excellent model for studying exocytosis. This gland synthesizes mucins that are packaged in SGs that sprout from the trans-Golgi network and then undergo a maturation process that involves homotypic fusion, condensation, and acidification. Finally, mature SGs are directed to the apical domain of the plasma membrane with which they fuse, releasing their content into the gland lumen. The exocyst is a hetero-octameric complex that participates in tethering of vesicles to the plasma membrane during constitutive exocytosis. By precise temperature-dependent gradual activation of the Gal4-UAS expression system, we have induced different levels of silencing of exocyst complex subunits, and identified three temporarily distinctive steps of the regulated exocytic pathway where the exocyst is critically required: SG biogenesis, SG maturation, and SG exocytosis. Our results shed light on previously unidentified functions of the exocyst along the exocytic pathway. We propose that the exocyst acts as a general tethering factor in various steps of this cellular process.
