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Intense Expression of GFP Gene in Gonads of Chicken Embryos by Transfecting Circulating Primordial Germ Cells in vitro and in vivo

  The present study was carried out to develop techniques for introducing exogenous DNA into primordial germ cells (PGCs) and expressing the introduced DNA efficiently in the gonads of developing chicken embryos. PGCs circulating in the bloodstream (stages 14-15) were transfected with GFP gene in vi... Full description

Journal Title: The Journal of Poultry Science 2007, Vol.44(4), p.416
Main Author: Naito, Mitsuru
Other Authors: Minematsu, Takeo , Harumi, Takashi , Kuwana, Takashi
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: ISSN: 13467395 ; E-ISSN: 13490486
Link: http://search.proquest.com/docview/1439293179/?pq-origsite=primo
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title: Intense Expression of GFP Gene in Gonads of Chicken Embryos by Transfecting Circulating Primordial Germ Cells in vitro and in vivo
format: Article
creator:
  • Naito, Mitsuru
  • Minematsu, Takeo
  • Harumi, Takashi
  • Kuwana, Takashi
subjects:
  • Gonads
  • Green Fluorescent Protein
  • Primordial Germ Cell
  • Transfection
ispartof: The Journal of Poultry Science, 2007, Vol.44(4), p.416
description:   The present study was carried out to develop techniques for introducing exogenous DNA into primordial germ cells (PGCs) and expressing the introduced DNA efficiently in the gonads of developing chicken embryos. PGCs circulating in the bloodstream (stages 14-15) were transfected with GFP gene in vitro or in vivo by lipofection or nucleofection. The manipulated PGCs successfully migrated to the germinal ridges and expressed GFP gene efficiently in the gonads of developing embryos. Intense GFP gene expression was observed in the gonads during the first 8 days following the transfection, during which period the sexual differentiation of gonads and germ cells (GCs) takes place. The GFP gene expression then gradually declined during subsequent embryonic development until hatching. When PGCs were transfected in vivo by lipofection with linearised plasmid DNA, GFP gene was detected in the gonads of 4.3% (19/442) of embryos examined at 20.5 days of culture, whereas less than 1% of embryos detected GFP gene in the gonads of embryos in which PGCs were transfected in vitro or with circular form plasmid DNA. In two of the embryos in which PGCs were transfected in vivo by lipofection with linearised plasmid DNA, GFP gene was expressed clearly in limited areas of the gonads of 20.5-day cultured embryos. The results obtained in this study suggest that the present in vitro and in vivo techniques for PGC manipulation provide a useful experimental system for studying gene functions in the sexual differentiation of gonads and GCs in early chicken embryos.
language: eng
source:
identifier: ISSN: 13467395 ; E-ISSN: 13490486
fulltext: fulltext
issn:
  • 13467395
  • 1346-7395
  • 13490486
  • 1349-0486
url: Link


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titleIntense Expression of GFP Gene in Gonads of Chicken Embryos by Transfecting Circulating Primordial Germ Cells in vitro and in vivo
creatorNaito, Mitsuru ; Minematsu, Takeo ; Harumi, Takashi ; Kuwana, Takashi
ispartofThe Journal of Poultry Science, 2007, Vol.44(4), p.416
identifierISSN: 13467395 ; E-ISSN: 13490486
description  The present study was carried out to develop techniques for introducing exogenous DNA into primordial germ cells (PGCs) and expressing the introduced DNA efficiently in the gonads of developing chicken embryos. PGCs circulating in the bloodstream (stages 14-15) were transfected with GFP gene in vitro or in vivo by lipofection or nucleofection. The manipulated PGCs successfully migrated to the germinal ridges and expressed GFP gene efficiently in the gonads of developing embryos. Intense GFP gene expression was observed in the gonads during the first 8 days following the transfection, during which period the sexual differentiation of gonads and germ cells (GCs) takes place. The GFP gene expression then gradually declined during subsequent embryonic development until hatching. When PGCs were transfected in vivo by lipofection with linearised plasmid DNA, GFP gene was detected in the gonads of 4.3% (19/442) of embryos examined at 20.5 days of culture, whereas less than 1% of embryos detected GFP gene in the gonads of embryos in which PGCs were transfected in vitro or with circular form plasmid DNA. In two of the embryos in which PGCs were transfected in vivo by lipofection with linearised plasmid DNA, GFP gene was expressed clearly in limited areas of the gonads of 20.5-day cultured embryos. The results obtained in this study suggest that the present in vitro and in vivo techniques for PGC manipulation provide a useful experimental system for studying gene functions in the sexual differentiation of gonads and GCs in early chicken embryos.
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description  The present study was carried out to develop techniques for introducing exogenous DNA into primordial germ cells (PGCs) and expressing the introduced DNA efficiently in the gonads of developing chicken embryos. PGCs circulating in the bloodstream (stages 14-15) were transfected with GFP gene in vitro or in vivo by lipofection or nucleofection. The manipulated PGCs successfully migrated to the germinal ridges and expressed GFP gene efficiently in the gonads of developing embryos. Intense GFP gene expression was observed in the gonads during the first 8 days following the transfection, during which period the sexual differentiation of gonads and germ cells (GCs) takes place. The GFP gene expression then gradually declined during subsequent embryonic development until hatching. When PGCs were transfected in vivo by lipofection with linearised plasmid DNA, GFP gene was detected in the gonads of 4.3% (19/442) of embryos examined at 20.5 days of culture, whereas less than 1% of embryos detected GFP gene in the gonads of embryos in which PGCs were transfected in vitro or with circular form plasmid DNA. In two of the embryos in which PGCs were transfected in vivo by lipofection with linearised plasmid DNA, GFP gene was expressed clearly in limited areas of the gonads of 20.5-day cultured embryos. The results obtained in this study suggest that the present in vitro and in vivo techniques for PGC manipulation provide a useful experimental system for studying gene functions in the sexual differentiation of gonads and GCs in early chicken embryos.
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abstract  The present study was carried out to develop techniques for introducing exogenous DNA into primordial germ cells (PGCs) and expressing the introduced DNA efficiently in the gonads of developing chicken embryos. PGCs circulating in the bloodstream (stages 14-15) were transfected with GFP gene in vitro or in vivo by lipofection or nucleofection. The manipulated PGCs successfully migrated to the germinal ridges and expressed GFP gene efficiently in the gonads of developing embryos. Intense GFP gene expression was observed in the gonads during the first 8 days following the transfection, during which period the sexual differentiation of gonads and germ cells (GCs) takes place. The GFP gene expression then gradually declined during subsequent embryonic development until hatching. When PGCs were transfected in vivo by lipofection with linearised plasmid DNA, GFP gene was detected in the gonads of 4.3% (19/442) of embryos examined at 20.5 days of culture, whereas less than 1% of embryos detected GFP gene in the gonads of embryos in which PGCs were transfected in vitro or with circular form plasmid DNA. In two of the embryos in which PGCs were transfected in vivo by lipofection with linearised plasmid DNA, GFP gene was expressed clearly in limited areas of the gonads of 20.5-day cultured embryos. The results obtained in this study suggest that the present in vitro and in vivo techniques for PGC manipulation provide a useful experimental system for studying gene functions in the sexual differentiation of gonads and GCs in early chicken embryos.
copIbaraki
pubJapan Science and Technology Agency
urlhttp://search.proquest.com/docview/1439293179/
pages416-425
doi10.2141/jpsa.44.416