title: Efficient gene delivery into murine ovarian cells by intraovarian injection of plasmid DNA and subsequent in vivo electroporation.
abstract: We describe the use of direct injection of circular plasmid DNA and subsequent in vivo electroporation (EP) for efficient gene delivery to the ovarian cells, including follicular cells and oocytes of mice. When Trypan blue (TB) was injected into the central portion of an ovary by a glass micropipette, rapid dispersion of TB to each preantral and antral follicle was observed. Injections of lacZ-expressing plasmid DNA and subsequent in vivo EP resulted in transfection of follicles with efficiencies ranging from 8-60%, together with cells in the thecal portion of the ovary. Of the lacZ-positive follicles, some oocytes were also positive for lacZ activity. These findings suggest that a solution introduced inside the ovary is rapidly dispersed to each follicle. With this technique, we expect great progress in genetic engineering in murine ovary.
authors.affiliation: The Institute of Medical Sciences, Tokai University, Bohseidai, Isehara, Kanagawa, Japan. masasato@is.icc.u-tokai.ac.jp
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sato
authors.forename: Maya
authors.initials: M
authors.lastname: Tanigawa
authors.forename: Natsuko
authors.initials: N
authors.lastname: Kikuchi
authors.forename: Shingo
authors.initials: S
authors.lastname: Nakamura
authors.forename: Minoru
authors.initials: M
authors.lastname: Kimura
mesh_terms: D000818:Animals; D018274:Electroporation / Q000379:methods*; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000008:administration & dosage / Q000378:metabolism; D007267:Injections / Q000379:methods; D051379:Mice; D010053:Ovary / Q000378:metabolism*; D010957:Plasmids / Q000008:administration & dosage / Q000378:metabolism*