Hypoxia modulates fibroblastic architecture, adhesion and migration: a role for HIF-1α in cofilin regulation and cytoplasmic actin distribution
Cells can adapt to hypoxia by various mechanisms. Yet, hypoxia-induced effects on the cytoskeleton-based cell architecture and functions are largely unknown. Here we present a comprehensive analysis of the architecture and function of L929 fibroblasts under hypoxic conditions (1% O2). Cells cultivat...
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| Veröffentlicht in: | PloS one 2013-07, Vol.8 (7), p.e69128-e69128 |
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| creator | Vogler, Melanie Vogel, Sabine Krull, Sabine Farhat, Katja Leisering, Pia Lutz, Susanne Wuertz, Christina M Katschinski, Dörthe M Zieseniss, Anke |
| description | Cells can adapt to hypoxia by various mechanisms. Yet, hypoxia-induced effects on the cytoskeleton-based cell architecture and functions are largely unknown. Here we present a comprehensive analysis of the architecture and function of L929 fibroblasts under hypoxic conditions (1% O2). Cells cultivated in hypoxia showed striking morphological differences as compared to cells cultivated under normoxic conditions (20% O2). These changes include an enlargement of cell area and volume, increased numbers of focal contacts and loss of cell polarization. Furthermore the β- and γ-actin distribution is greatly altered. These hypoxic adjustments are associated with enhanced cell spreading and a decline of cell motility in wound closure and single cell motility assays. As the hypoxia-inducible factor-1α (HIF-1α) is stabilised in hypoxia and plays a pivotal role in the transcriptional response to changes in oxygen availability we used an shRNA-approach to examine the role of HIF-1α in cytoskeleton-related architecture and functions. We show that the observed increase in cell area, actin filament rearrangement, decrease of single cell migration in hypoxia and the maintenance of p-cofilin levels is dependent on HIF-1α stabilisation. |
| doi_str_mv | 10.1371/journal.pone.0069128 |
| format | Article |
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Yet, hypoxia-induced effects on the cytoskeleton-based cell architecture and functions are largely unknown. Here we present a comprehensive analysis of the architecture and function of L929 fibroblasts under hypoxic conditions (1% O2). Cells cultivated in hypoxia showed striking morphological differences as compared to cells cultivated under normoxic conditions (20% O2). These changes include an enlargement of cell area and volume, increased numbers of focal contacts and loss of cell polarization. Furthermore the β- and γ-actin distribution is greatly altered. These hypoxic adjustments are associated with enhanced cell spreading and a decline of cell motility in wound closure and single cell motility assays. As the hypoxia-inducible factor-1α (HIF-1α) is stabilised in hypoxia and plays a pivotal role in the transcriptional response to changes in oxygen availability we used an shRNA-approach to examine the role of HIF-1α in cytoskeleton-related architecture and functions. We show that the observed increase in cell area, actin filament rearrangement, decrease of single cell migration in hypoxia and the maintenance of p-cofilin levels is dependent on HIF-1α stabilisation.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0069128</identifier><identifier>PMID: 23874890</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Actin ; Actin Depolymerizing Factors - metabolism ; Actins - metabolism ; Analysis of Variance ; Animals ; Architecture ; Biology ; Cell adhesion & migration ; Cell Adhesion - physiology ; Cell Hypoxia - physiology ; Cell Line, Tumor ; Cell migration ; Cell Movement - physiology ; Cell Size ; Cell spreading ; Cofilin ; Cytoplasm - metabolism ; Cytoskeleton ; Enlargement ; Fibroblasts ; Fibroblasts - cytology ; Fibroblasts - physiology ; Flow Cytometry ; Fluorescence ; Gene Knockdown Techniques ; Hypoxia ; Hypoxia-Inducible Factor 1, alpha Subunit - metabolism ; Hypoxia-inducible factors ; Immunoblotting ; Kinases ; Medicine ; Mice ; Morphology ; Motility ; Oxygen ; Phosphorylation ; Physics ; Physiology ; Proteins ; Pulmonary arteries ; Rodents ; Studies ; Transcription ; Wound healing ; Wounds</subject><ispartof>PloS one, 2013-07, Vol.8 (7), p.e69128-e69128</ispartof><rights>2013 Vogler et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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Yet, hypoxia-induced effects on the cytoskeleton-based cell architecture and functions are largely unknown. Here we present a comprehensive analysis of the architecture and function of L929 fibroblasts under hypoxic conditions (1% O2). Cells cultivated in hypoxia showed striking morphological differences as compared to cells cultivated under normoxic conditions (20% O2). These changes include an enlargement of cell area and volume, increased numbers of focal contacts and loss of cell polarization. Furthermore the β- and γ-actin distribution is greatly altered. These hypoxic adjustments are associated with enhanced cell spreading and a decline of cell motility in wound closure and single cell motility assays. As the hypoxia-inducible factor-1α (HIF-1α) is stabilised in hypoxia and plays a pivotal role in the transcriptional response to changes in oxygen availability we used an shRNA-approach to examine the role of HIF-1α in cytoskeleton-related architecture and functions. We show that the observed increase in cell area, actin filament rearrangement, decrease of single cell migration in hypoxia and the maintenance of p-cofilin levels is dependent on HIF-1α stabilisation.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>23874890</pmid><doi>10.1371/journal.pone.0069128</doi><oa>free_for_read</oa></addata></record> |
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| subjects | Actin Actin Depolymerizing Factors - metabolism Actins - metabolism Analysis of Variance Animals Architecture Biology Cell adhesion & migration Cell Adhesion - physiology Cell Hypoxia - physiology Cell Line, Tumor Cell migration Cell Movement - physiology Cell Size Cell spreading Cofilin Cytoplasm - metabolism Cytoskeleton Enlargement Fibroblasts Fibroblasts - cytology Fibroblasts - physiology Flow Cytometry Fluorescence Gene Knockdown Techniques Hypoxia Hypoxia-Inducible Factor 1, alpha Subunit - metabolism Hypoxia-inducible factors Immunoblotting Kinases Medicine Mice Morphology Motility Oxygen Phosphorylation Physics Physiology Proteins Pulmonary arteries Rodents Studies Transcription Wound healing Wounds |
| title | Hypoxia modulates fibroblastic architecture, adhesion and migration: a role for HIF-1α in cofilin regulation and cytoplasmic actin distribution |
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