title: Identification and characterization of an oocyte factor required for development of porcine nuclear transfer embryos.
abstract: Nuclear reprogramming of differentiated cells can be induced by oocyte factors. Despite numerous attempts, these factors and mechanisms responsible for successful reprogramming remain elusive. Here, we identify one such factor, necessary for the development of nuclear transfer embryos, using porcine oocyte extracts in which some reprogramming events are recapitulated. After incubating somatic nuclei in oocyte extracts from the metaphase II stage, the oocyte proteins that were specifically and abundantly incorporated into the nuclei were identified by mass spectrometry. Among 25 identified proteins, we especially focused on a multifunctional protein, DJ-1. DJ-1 is present at a high concentration in oocytes from the germinal vesicle stage until embryos at the four-cell stage. Inhibition of DJ-1 function compromises the development of nuclear transfer embryos but not that of fertilized embryos. Microarray analysis of nuclear transfer embryos in which DJ-1 function is inhibited shows perturbed expression of P53 pathway components. In addition, embryonic arrest of nuclear transfer embryos injected with anti-DJ-1 antibody is rescued by P53 inhibition. We conclude that DJ-1 is an oocyte factor that is required for development of nuclear transfer embryos. This study presents a means for identifying natural reprogramming factors in mammalian oocytes and a unique insight into the mechanisms underlying reprogramming by nuclear transfer.
authors.affiliation: Laboratory of Reproductive Biology, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Kyoto 606-8502, Japan. kei_m@kais.kyoto-u.ac.jp
authors.forename: Kei
authors.initials: K
authors.lastname: Miyamoto
authors.forename: Kouhei
authors.initials: K
authors.lastname: Nagai
authors.forename: Naoya
authors.initials: N
authors.lastname: Kitamura
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Nishikawa
authors.forename: Haruka
authors.initials: H
authors.lastname: Ikegami
authors.forename: Nguyen T
authors.initials: NT
authors.lastname: Binh
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tsukamoto
authors.forename: Mai
authors.initials: M
authors.lastname: Matsumoto
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Tsukiyama
authors.forename: Naojiro
authors.initials: N
authors.lastname: Minami
authors.forename: Masayasu
authors.initials: M
authors.lastname: Yamada
authors.forename: Hiroyoshi
authors.initials: H
authors.lastname: Ariga
authors.forename: Masashi
authors.initials: M
authors.lastname: Miyake
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Kawarasaki
authors.forename: Kazuya
authors.initials: K
authors.lastname: Matsumoto
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Imai
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D054337:Cell Dedifferentiation / Q000187:drug effects*; D005260:Female; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000494:pharmacology*; D008677:Metaphase / Q000502:physiology; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism*; D013552:Swine; D016159:Tumor Suppressor Protein p53 / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D016159:Tumor Suppressor Protein p53