title: Establishment of oct4:egfp transgenic and oct4:egfp /β-actin:DsRed double transgenic medaka lines.
abstract: As a model to examine cellular multipotency in fish, we established a medaka transgenic (Tg) Tru.oct4:egfp line carrying the green fluorescence protein (GFP) cDNA under control of the Takifugu rubripes oct4 promoter. In this Tg line, GFP could be used to examine both maternal and zygotic oct4 expression during embryogenesis. In addition, while adult Tg fish did not express GFP in any somatic cells, activation of GFP expression was initiated in regenerating fins after amputation. In vitro, some of the cell populations that migrated from fin explants expressed GFP, implying that GFP could be used to monitor oct4 expression in both embryos and in regenerating tissues in the Tru.oct4:egfp Tg line. Next, crossing with β-actin:DsRed Tg line in which all cells emit red fluorescence by expression of red fluorescent protein (RFP) under the β-actin promoter, we prepared a Tru.oct4:egfp /β-actin:DsRed double Tg line. In the double Tg line, early embryonic cells were +GFP/+RFP double positive. In vitro fin cell culture, a small number of +GFP/+RFP double positive cells could be discriminated from other -GFP/+RFP cells. Thus, when transplanted into wild-type medaka, this double Tg line can be used to trace the fate of the transplanted cells using RFP fluorescence after the loss of GFP expression.
authors.affiliation: Laboratory of Marine Life Science and Genetics, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan.
authors.forename: Shinpei
authors.initials: S
authors.lastname: Yokota
authors.affiliation: Laboratory of Marine Life Science and Genetics, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan.
authors.forename: Rinta
authors.initials: R
authors.lastname: Matsuno
authors.affiliation: Laboratory of Marine Life Science and Genetics, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kato
authors.affiliation: Bioscience and Biotechnology Center, Nagoya University, Nagoya, 464-8601, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Hashimoto
authors.affiliation: Division of Applied Bioscience, Graduate School of Agriculture, Kyoto University, Kyoto, 606-8502, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Kinoshita
authors.affiliation: Laboratory of Marine Life Science and Genetics, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan.
authors.forename: Hayato
authors.initials: H
authors.lastname: Yokoi
authors.affiliation: Laboratory of Marine Life Science and Genetics, Graduate School of Agricultural Science, Tohoku University, Sendai, 981-8555, Japan. suzukitr@bios.tohoku.ac.jp.
authors.forename: Tohru
authors.initials: T
authors.lastname: Suzuki
mesh_terms: D000199:Actins / Q000235:genetics* / Q000378:metabolism; D058500:Animal Fins / Q000378:metabolism / Q000502:physiology; D000818:Animals; D030801:Animals, Genetically Modified; D017690:Cell Transplantation / Q000662:veterinary; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000235:genetics* / Q000378:metabolism; D009990:Oryzias / Q000235:genetics* / Q000378:metabolism; D012038:Regeneration; D000097573:Red Fluorescent Protein
chemical_list: D000199:Actins; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D050814:Octamer Transcription Factor-3; D000097573:Red Fluorescent Protein
keywords: Embryo; Fin regeneration; GFP; Medaka; RFP; Transgenic fish; oct4