title: Effects of downregulating GLIS1 transcript on preimplantation development and gene expression of bovine embryos.
abstract: Krüppel-like protein Gli-similar 1 (GLIS1) is known as a direct reprogramming factor for the generation of induced pluripotent stem cells. The objective of this study was to investigate the role of GLIS1 in the preimplantation development of bovine embryos. GLIS1 transcripts in in vitro-matured oocytes and 1-cell to 4-cell stage embryos were detected, but they were either absent or at trace levels at the 8-cell to blastocyst stages. We attempted GLIS1 downregulation of bovine early embryos by RNA interference and evaluated developmental competency and gene transcripts, which are involved in zygotic gene activation (ZGA) in GLIS1-downregulated embryos. Injection of specific siRNA resulted in a distinct decrease in GLIS1 transcript in bovine embryos at the 4-cell stage. Although the bovine embryos injected with GLIS1-siRNA could develop to the 16-cell stage, these embryos had difficulty in developing beyond the 32-cell stage. Gene transcripts of PDHA1 and HSPA8, which are transcribed after ZGA, showed lower level in GLIS1 downregulated embryos. It is possible that GLIS1-downregulated embryos fail to initiate ZGA. Our results indicated that GLIS1 is an important factor for the preimplantation development of bovine embryos.
authors.affiliation: Faculty of Agriculture, Iwate University, Iwate 020-8550, Japan.
authors.forename: Kazuki
authors.initials: K
authors.lastname: Takahashi
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Sakurai
authors.forename: Natsuko
authors.initials: N
authors.lastname: Emura
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Hashizume
authors.forename: Ken
authors.initials: K
authors.lastname: Sawai
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D001757:Blastomeres / Q000166:cytology / Q000378:metabolism*; D002417:Cattle; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D004477:Ectogenesis; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D018507:Gene Expression Regulation, Developmental; D050883:HSC70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D059471:In Vitro Oocyte Maturation Techniques / Q000662:veterinary; D008845:Microinjections / Q000662:veterinary; D009028:Morula / Q000166:cytology / Q000378:metabolism; D009865:Oocytes / Q000166:cytology / Q000378:metabolism*; D030481:Pyruvate Dehydrogenase (Lipoamide) / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D015053:Zygote / Q000166:cytology / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; D050883:HSC70 Heat-Shock Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014157:Transcription Factors; D030481:Pyruvate Dehydrogenase (Lipoamide); C065640:pyruvate dehydrogenase E1alpha subunit