Effects of downregulating GLIS1 transcript on preimplantation development and gene expression of bovine embryos

abstract: ...Krppel-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.

...J Reprod Dev J. Reprod. Dev JRD The Journal of Reproduction and Development 0916-8818 1348-4400 The Society for Reproduction and Development 26074126 4623141 2015-029 10.1262/jrd.2015-029 Original Article Effects of downregulating GLIS1 transcript on preimplantation development and gene expression of bovine embryos TAKAHASHI Kazuki 1 SAKURAI Nobuyuki 2 EMURA Natsuko 1 HASHIZUME Tsutomu 1 2 SAWAI Ken 1 2 1) Faculty of Agriculture, Iwate University, Iwate 020-8550, Japan 2) The United Graduate School of Agricultural Sciences, Iwate University, Iwate 020-8550, Japan Correspondences: K Sawai (e-mail: kensawai@iwate-u.ac.jp ) 15 6 2015 10 2015 61 5 369 374 05 3 2015 01 5 2015 2015 Society for Reproduction and Development 2015 This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. Krppel-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. Bovine embryo Early development Gene expression GLIS1 RNA interference Although the procedures for in vitro production (IVP) of domestic animal embryos such as in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) have improved, the efficiencies of embryo development and offspring production after embryo transfer are not high. In particular, neonatal and postnatal

... to an embryonic state [ 12 , 13 ]. However, the reprogramming factor(s) in oocytes have not yet been determined, and limited information concerning the mechanism of nuclear reprogramming in oocytes is available. The Krppel-like protein Gli-similar 1 (GLIS1) constitutes a subfamily of Krppel-like zinc finger transcription factors that are closely related to the Gli family [ 14 , 15 , 16 ]. Recently, GLIS1 has been shown to be a direct reprogramming factor for somatic cell nuclei, in that, GLIS1 markedly enhances the generation of induced pluripotent stem cells (iPSCs) from both mouse and human fibroblasts when it is expressed along with Pou5f1 ( OCT3/4 ), SOX2 and KLF4 [ 17 ]. GLIS1 is able to replace oncogenic MYC , resulting in decreased tumorigenicity as well as improved safety and efficiency of iPSCs production [ 17 ]. Notably, Glis1 transcript is enriched in unfertilized oocytes and 1-cell stage embryos in mice [ 17 ]. Furthermore, Glis1 was reported to be both temporally and spatially

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Gene: GLIS1 - GLIS family zinc finger 1 - (NCBI)

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