Rapidly migrating zebrafish keratocytes and their interactions with the physical environment
Abstract/Contents
- Abstract
- Skin wound healing provides essential protection to multicellular organisms. To successfully heal, skin cells need to migrate to cover the wound bed. For mammals this migration occurs during many days and is hidden beneath the scab; therefore, we leverage the larval zebrafish to image their rapid wound healing (duration of < 30 minutes). Zebrafish basal epidermal cells, or keratocytes, migrate towards wounds using the actin cytoskeleton. Keratocyte migration can additionally be studied in isolated cell culture conditions, allowing experimental separation of the many physical factors present at wound sites. Here, we dissect the complex interplay between migrating skin cells and their environment using both isolated keratocytes in various micro-engineered environments and in vivo keratocytes in the skin of larval zebrafish. First, we investigated the speed of isolated keratocytes on nano-topographical substrate features, which have relevance to biomaterial design. Nanopillars caused slower cell speed, in association with actin accumulation on the pillars. Aberrant actin polymerization may effectively increase cell-substrate adhesion, leading to the observed rear retraction defect. This study demonstrates that keratocytes adjust their molecular- and cell-scale behaviors in response to non-molecular changes in the environment. We next examined isolated keratocytes in multiple two-dimensionally confining environments, which mimic the flat, thin shape of the zebrafish epidermis. Confined isolated cells recapitulate the shape of their in vivo counterparts but migrate more slowly than unconfined cells. We observed that cell speed reduction is associated with cell shrinkage following both confinement and hypertonic shock, potentially due to cytoplasmic crowding. We speculate that this volume-speed relationship may promote rapid migration following injury in freshwater, which causes cell swelling. Finally, we returned to in vivo wound healing to explore the physical implications of freshwater injury. We discovered a novel fluid force during late stages of wound healing, which causes fracture events between adjacent cells. Keratocytes respond by intaking large volumes of excess fluid. Overall, these studies highlight the need to consider molecularly simple contributors, including geometry and water, because of their surprising effects on molecular networks, cell behavior, and ultimately healing efficacy.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2022; ©2022 |
| Publication date | 2022; 2022 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Labuz, Ellen Claire |
|---|---|
| Degree supervisor | Dunn, Alexander Robert |
| Thesis advisor | Dunn, Alexander Robert |
| Thesis advisor | Wang, Bo, (Artificial intelligence scientist) |
| Thesis advisor | Zuchero, J |
| Degree committee member | Wang, Bo, (Artificial intelligence scientist) |
| Degree committee member | Zuchero, J |
| Associated with | Stanford University, Biophysics Program |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Ellen Claire Labuz. |
|---|---|
| Note | Submitted to the Biophysics Program. |
| Thesis | Thesis Ph.D. Stanford University 2022. |
| Location | https://purl.stanford.edu/jg737jz6969 |
Access conditions
- Copyright
- © 2022 by Ellen Claire Labuz
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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