A murine living skin equivalent amenable to live-cell imaging: analysis of the roles of connexins in the epidermis.
Three-dimensional (3D) organotypic models are increasingly used to study the aspects of epidermal organisation and cutaneous wound-healing events. However, these are largely dependent on laborious histological analysis and immunohistochemical approaches. Despite the large resource of transgenic and knockout mice harboring mutations relevant to skin disorders, few organotypic mouse skin models are available. We have developed a versatile in vitro 3D organotypic mouse skin equivalent that reflects epidermal organisation in vivo. The system is optically transparent and ideally suited to real-time analysis using a variety of integrated in situ imaging techniques. As a paradigm for coordination of cellular events, the epidermal gap junction network was investigated and the model displayed predominant connexin 43 (Cx43) expression in basal proliferating cells and Cx26 and Cx30 expression in differentiated keratinocytes. We show that attenuation of Cx43-mediated communication by a Cx mimetic peptide enhanced wound closure rates in keratinocyte monocultures and in the living skin equivalent system, emphasising the utility of the model to systematically unravel the molecular mechanisms underlying epidermal morphogenesis, assess promising therapeutic strategies, and reduce animal experimentation. Furthermore, we visualise epidermal regeneration following injury in real time, thereby facilitating avenues to explore distinctive modes of wound re-epithelialisation in a non-invasive manner.
Kandyba
Eve E
EE
Department of Biological and Biomedical Sciences, School of Life Sciences, Glasgow Caledonian University, Glasgow, UK.
Hodgins
Malcolm B
MB
Martin
Patricia E
PE
D000818:Animals; D000072259:Connexin 26; D000075302:Connexin 30; D018031:Connexin 43 / Q000032:analysis / Q000502:physiology; D017630:Connexins / Q000032:analysis / Q000502:physiology*; D000078404:Epidermal Cells; D004817:Epidermis / Q000187:drug effects / Q000502:physiology*; D017629:Gap Junctions; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D051379:Mice; D009924:Organ Culture Techniques / Q000379:methods*; D012038:Regeneration; D012867:Skin / Q000166:cytology / Q000187:drug effects; D014945:Wound Healing / Q000187:drug effects
D000075302:Connexin 30; D018031:Connexin 43; D017630:Connexins; C100629:Gjb6 protein, mouse; D000072259:Connexin 26