title: Three-dimensional morphogenesis of MDCK cells induced by cellular contractile forces on a viscous substrate.
abstract: Substrate physical properties are essential for many physiological events such as embryonic development and 3D tissue formation. Physical properties of the extracellular matrix such as viscoelasticity and geometrical constraints are understood as factors that affect cell behaviour. In this study, we focused on the relationship between epithelial cell 3D morphogenesis and the substrate viscosity. We observed that Madin-Darby Canine Kidney (MDCK) cells formed 3D structures on a viscous substrate (Matrigel). The structures appear as a tulip hat. We then changed the substrate viscosity by genipin (GP) treatment. GP is a cross-linker of amino groups. Cells cultured on GP-treated-matrigel changed their 3D morphology in a substrate viscosity-dependent manner. Furthermore, to elucidate the spatial distribution of the cellular contractile force, localization of mono-phosphorylated and di-phosphorylated myosin regulatory light chain (P-MRLCs) was visualized by immunofluorescence. P-MRLCs localized along the periphery of epithelial sheets. Treatment with Y-27632, a Rho-kinase inhibitor, blocked the P-MRLCs localization at the edge of epithelial sheets and halted 3D morphogenesis. Our results indicate that the substrate viscosity, the substrate deformation, and the cellular contractile forces induced by P-MRLCs play crucial roles in 3D morphogenesis.
authors.affiliation: Transdisciplinary Life Science Course, Faculty of Advanced Life Science, Hokkaido University, N10-W8, Kita-ku, Sapporo 060-0810, Japan.
authors.forename: Misako
authors.initials: M
authors.lastname: Imai
authors.affiliation: Transdisciplinary Life Science Course, Faculty of Advanced Life Science, Hokkaido University, N10-W8, Kita-ku, Sapporo 060-0810, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Furusawa
authors.affiliation: Transdisciplinary Life Science Course, Faculty of Advanced Life Science, Hokkaido University, N10-W8, Kita-ku, Sapporo 060-0810, Japan.
authors.forename: Takeomi
authors.initials: T
authors.lastname: Mizutani
authors.affiliation: Transdisciplinary Life Science Course, Faculty of Advanced Life Science, Hokkaido University, N10-W8, Kita-ku, Sapporo 060-0810, Japan.
authors.forename: Kazushige
authors.initials: K
authors.lastname: Kawabata
authors.affiliation: Transdisciplinary Life Science Course, Faculty of Advanced Life Science, Hokkaido University, N10-W8, Kita-ku, Sapporo 060-0810, Japan.|Research Center for Cooperative Projects, Hokkaido University Graduate School of Medicine, Sapporo, 060-8638, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Haga
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D003094:Collagen; D004285:Dogs; D004338:Drug Combinations; D005109:Extracellular Matrix; D007797:Laminin; D061985:Madin Darby Canine Kidney Cells / Q000166:cytology* / Q000502:physiology*; D018994:Myosin Light Chains / Q000378:metabolism; D010766:Phosphorylation; D011509:Proteoglycans; D018874:Spheroids, Cellular; D014783:Viscosity
chemical_list: D004338:Drug Combinations; D007797:Laminin; D018994:Myosin Light Chains; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen