Journal article
Mouse Keratinocytes Without Keratin Intermediate Filaments Demonstrate Substrate Stiffness Dependent Behaviors
Cellular and molecular bioengineering, Vol.11(3), pp.163-174
06/01/2018
DOI: 10.1007/s12195-018-0526-y
PMCID: PMC6816603
PMID: 31719883
Abstract
Introduction: Traditionally thought to serve active vs. passive mechanical functions, respectively, a growing body of evidence suggests that actin microfilament and keratin intermediate filament (IF) networks, together with their associated cell-cell and cell-matrix anchoring junctions, may have a large degree of functional interdependence. Therefore, we hypothesized that the loss of keratin IFs in a knockout mouse keratinocyte model would affect the kinematics of colony formation, i.e., the spatiotemporal process by which individual cells join to form colonies and eventually a nascent epithelial sheet.
Methods: Time-lapse imaging and deformation tracking microscopy was used to observe colony formation for both wild type (WT) and keratin-deficient knockout (KO) mouse keratinocytes over 24 h. Cells were cultured under high calcium conditions on collagen-coated substrates with nominal stiffnesses of ~ 1.2 kPa (soft) and 24 kPa (stiff). Immunofluorescent staining of actin and selected adhesion proteins was also performed.
Results: The absence of keratin IFs markedly affected cell morphology, spread area, and cytoskeleton and adhesion protein organization on both soft and stiff substrates. Strikingly, an absence of keratin IFs also significantly reduced the ability of mouse keratinocytes to mechanically deform the soft substrate. Furthermore, KO cells formed colonies more efficiently on stiff vs. soft substrates, a behavior opposite to that observed for WT keratinocytes.
Conclusions: Collectively, these data are strongly supportive of the idea that an interdependence between actin microfilaments and keratin IFs does exist, while further suggesting that keratin IFs may represent an important and under-recognized component of keratinocyte mechanosensation and the force generation apparatus.
Details
- Title: Subtitle
- Mouse Keratinocytes Without Keratin Intermediate Filaments Demonstrate Substrate Stiffness Dependent Behaviors
- Creators
- Hoda Zarkoob - 5629 Seamans Center, Iowa City, IA USASathivel Chinnathambi - 5629 Seamans Center, Iowa City, IA USASpencer A Halberg - 5629 Seamans Center, Iowa City, IA USAJohn C Selby - Iowa City, IA USAThomas M Magin - 04103 Leipzig, GermanyE. A Sander - 5629 Seamans Center, Iowa City, IA USA
- Resource Type
- Journal article
- Publication Details
- Cellular and molecular bioengineering, Vol.11(3), pp.163-174
- DOI
- 10.1007/s12195-018-0526-y
- PMID
- 31719883
- PMCID
- PMC6816603
- NLM abbreviation
- Cell Mol Bioeng
- ISSN
- 1865-5025
- eISSN
- 1865-5033
- Publisher
- Springer US; New York
- Grant note
- #14-4384; #18-5045 / ; ; 1452728 / ; MA1316-15; MA1316-17; MA1316-19; INST 268/230-1 / ; MA1316-21 / ;
- Language
- English
- Date published
- 06/01/2018
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Dermatology; Orthopedics and Rehabilitation; Craniofacial Anomalies Research Center; Chemical and Biochemical Engineering
- Record Identifier
- 9984025694202771
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