title: In vivo imaging of tight junctions using claudin-EGFP transgenic medaka.
abstract: Tight junctions (TJs) function as a physiological barrier between epithelial and endothelial sheets by restricting the diffusion of fluid through the intercellular space. Recent morphological studies associated with TJs have revealed that the TJ is a dynamic rather than a static structure; indeed, several in vitro studies indicate that proper TJ function requires dynamic TJ behavior. Direct observation of the dynamic behavior of TJs is necessary to understand the essential roles of TJs in physiological contexts, such as during embryogenesis and metastasis. Here we describe a protocol for the generation of transgenic medaka (Oryzias latipes) that express claudin-EGFP. This fluorescent fusion protein enables real-time imaging of TJs in the living embryo. Claudin-EGFP transgenic medaka will be a useful tool to screen for mutations and for small molecules affecting cell-cell adhesion.
authors: Department of Genetics and Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan. t-miyamoto@hiroshima-u.ac.jp
authors: Tatsuo
authors: T
authors: Miyamoto
authors: Mikio
authors: M
authors: Furuse
authors: Makoto
authors: M
authors: Furutani-Seiki
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D057167:Claudins / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D008845:Microinjections / Q000379:methods; D008853:Microscopy / Q000379:methods; D009990:Oryzias / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D012685:Sepharose; D019108:Tight Junctions / Q000648:ultrastructure*; D014162:Transfection / Q000379:methods
chemical_list: D057167:Claudins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D012685:Sepharose