title: Zebrafish keratocyte explant cultures as a wound healing model system: differential gene expression & morphological changes support epithelial-mesenchymal transition.
abstract: The control of collective cell migration of zebrafish keratocyte sheets in explant culture is of interest for cell migration and epithelial wound healing and depends on the gene expression profile. In a zebrafish genome array, ∼17.5% of the probe sets were differentially expressed greater than two-fold (p≤0.003) between 1 and 7 days of explant culture. Among the differentially expressed genes were a variety of wound healing-related genes and many of the biomarkers for epithelial-mesenchymal transition (EMT), including a switch from keratin and E-cadherin to vimentin and N-cadherin expression and several EMT-related transcription factors were found to be differentially expressed. Supporting evidence for EMT is seen in both morphological change and rearrangement of the actin cytoskeleton and in expression of cadherins during explant culture with a visible disassembly of the cell sheet. TGFβ1 and TNFα expression were analyzed by qPCR at various time points and peak differential expression of both cytokines occurred at 3 days, indicating that the EMT process is ongoing under conditions routinely used in the study of fish keratocyte motility. These data establish that an EMT process is occurring during zebrafish keratocyte explant culture and support the use of this system as a wound healing model.
authors.affiliation: Biomedical Sciences, College of Health Sciences, Midwestern University, AZ, United States.
authors.forename: Timothy M
authors.initials: TM
authors.lastname: McDonald
authors.affiliation: Biomedical Sciences, College of Health Sciences, Midwestern University, AZ, United States.
authors.forename: Agnes S
authors.initials: AS
authors.lastname: Pascual
authors.affiliation: Microbiology & Immunology, Arizona College of Osteopathic Medicine, Midwestern University, AZ, United States.
authors.forename: Chandana K
authors.initials: CK
authors.lastname: Uppalapati
authors.affiliation: Biomedical Sciences, College of Health Sciences, Midwestern University, AZ, United States.
authors.forename: Kimbal E
authors.initials: KE
authors.lastname: Cooper
authors.affiliation: Microbiology & Immunology, Arizona College of Osteopathic Medicine, Midwestern University, AZ, United States.
authors.forename: Kathryn J
authors.initials: KJ
authors.lastname: Leyva
authors.affiliation: Biomedical Sciences, College of Health Sciences, Midwestern University, AZ, United States. Electronic address: ehullx@midwestern.edu.
authors.forename: Elizabeth E
authors.initials: EE
authors.lastname: Hull
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D002465:Cell Movement; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D020869:Gene Expression Profiling; D066298:In Vitro Techniques; D007633:Keratins / Q000235:genetics / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D014158:Transcription, Genetic; D016211:Transforming Growth Factor alpha / Q000235:genetics / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism; D014746:Vimentin / Q000235:genetics / Q000378:metabolism; D014945:Wound Healing / Q000235:genetics*; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D015820:Cadherins; D016211:Transforming Growth Factor alpha; D053773:Transforming Growth Factor beta1; D014746:Vimentin; D029961:Zebrafish Proteins; D007633:Keratins