title: Myosin II activity is required for functional leading-edge cells and closure of epidermal sheets in fish skin ex vivo.
abstract: Re-epithelialization in skin wound healing is a process in which epidermal sheets grow and close the wound. Although the actin-myosin system is thought to have a pivotal role in re-epithelialization, its role is not clear. In fish skin, re-epithelialization occurs around 500 μm/h and is 50 times faster than in mammalian skin. We had previously reported that leading-edge cells of the epidermal outgrowth have both polarized large lamellipodia and "purse string"-like actin filament cables in the scale-skin culture system of medaka fish, Oryzias latipes (Cell Tissue Res, 2007). The actin purse-string (APS) is a supracellular contractile machinery in which adherens junctions (AJs) link intracellular myosin II-including actin cables between neighboring cells. In this study, we developed a modified "face-to-face" scale-skin culture system as an ex vivo model to study epidermal wound healing, and examined the role of the actin-myosin system in the rapid re-epithelialization using a myosin II ATPase inhibitor, blebbistatin. A low level of blebbistatin suppressed the formation of APS and induced the dissociation of keratocytes from the leading edge without attenuating the growth of the epidermal sheet or the migration rate of solitary keratocytes. AJs in the superficial layer showed no obvious changes elicited by blebbistatin. However, two epidermal sheets without APSs did not make a closure with each other, which was confirmed by inhibiting the connecting AJs between the superficial layers. These results suggest that myosin II activity is required for functional leading-edge cells and for epidermal closure.
authors.affiliation: Department of Biomolecular Science, Toho University, Miyama, Funabashi, Chiba, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Morita
authors.forename: Akiko
authors.initials: A
authors.lastname: Tsuchiya
authors.forename: Masazumi
authors.initials: M
authors.lastname: Sugimoto
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002465:Cell Movement / Q000187:drug effects; D004817:Epidermis / Q000187:drug effects / Q000254:growth & development / Q000473:pathology*; D006576:Heterocyclic Compounds, 4 or More Rings / Q000494:pharmacology; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D024462:Myosin Type II / Q000378:metabolism*; D009924:Organ Culture Techniques; D009990:Oryzias; D010641:Phenotype; D011554:Pseudopodia / Q000187:drug effects / Q000378:metabolism*; D013997:Time Factors; D016596:Vinculin / Q000378:metabolism; D014945:Wound Healing / Q000187:drug effects
chemical_list: D000199:Actins; D006576:Heterocyclic Compounds, 4 or More Rings; D016596:Vinculin; C472645:blebbistatin; D024462:Myosin Type II