title: Generation of a transgenic medaka (Oryzias latipes) strain for visualization of nuclear dynamics in early developmental stages.
abstract: In this study, we verified nuclear transport activity of an artificial nuclear localization signal (aNLS) in medaka fish (Oryzias latipes). We generated a transgenic medaka strain expresses the aNLS tagged enhanced green fluorescent protein (EGFP) driven by a medaka beta-actin promoter. The aNLS-EGFP was accumulated in the nuclei of somatic tissues and yolk nuclei of oocytes, but undetectable in the spermatozoa. The fluorescent signal was observed from immediately after fertilization by a maternal contribution. Furthermore, male and female pronuclei were visualized in fertilized eggs, and nuclear dynamics of pronuclear fusion and subsequent cleavage were captured by time-lapse imaging. In contrast, SV40NLS exhibited no activity of nuclear transport in early embryos. In conclusion, the aNLS possesses a strong nuclear localization activity and is a useful probe for fluorescent observation of the pronuclei and nuclei in early developmental stage of medaka.
authors.affiliation: Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake-cho, Sakyo-ku, Kyoto, 606-8502, Japan.
authors.forename: Takanobu
authors.initials: T
authors.lastname: Inoue
authors.affiliation: Department of Growth Regulation, Institute for Frontier Medical Sciences, Kyoto University, Shogo-in Kawahara-cho 53, Sakyo-ku, Kyoto, 606-8507, Japan.
authors.forename: Atsuo
authors.initials: A
authors.lastname: Iida
authors.affiliation: Department of Intelligence Science and Technology, Graduate School of Informatics, Kyoto University, Yoshida-Honmachi, Sakyo-ku, Kyoto, 606-8501, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Maegawa
authors.affiliation: Department of Growth Regulation, Institute for Frontier Medical Sciences, Kyoto University, Shogo-in Kawahara-cho 53, Sakyo-ku, Kyoto, 606-8507, Japan.
authors.forename: Atsuko
authors.initials: A
authors.lastname: Sehara-Fujisawa
authors.affiliation: Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake-cho, Sakyo-ku, Kyoto, 606-8502, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Kinoshita
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology; D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology / Q000235:genetics; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D005260:Female; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D008856:Microscopy, Fluorescence / Q000379:methods; D009990:Oryzias / Q000196:embryology / Q000235:genetics
chemical_list: D049452:Green Fluorescent Proteins
keywords: medaka; nuclear localization signal; pronuclear fusion