Effects of downregulating GLIS1 transcript on preimplantation development and gene expression of bovine embryos.
Effects of downregulating GLIS1 transcript on preimplantation development and gene expression of bovine embryos.
复制标题
DOI：
10.1262/jrd.2015-029
复制
发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Sawai K
中科院分区：
文献类型：
--
作者：
Takahashi K;Sakurai N;Emura N;Hashizume T;Sawai K
KRüppel样蛋白Gli-similar 1(GLIS1)被认为是诱导多能干细胞产生的直接重编程因子。本研究旨在探讨GLIS1在牛胚胎着床前发育中的作用。在体外成熟卵母细胞和1-细胞-4-细胞期胚胎中检测到GLIS1转录本，但在8-细胞期-囊胚期GLIS1基因不存在或处于微量水平。我们尝试利用RNA干扰技术对牛早期胚胎进行GLIS1的下调，并评估了与GLIS1下调的胚胎合子基因激活(ZGA)相关的发育能力和基因转录产物。注射特异性siRNA后，4-细胞期牛胚胎GLIS1转录水平明显降低。虽然注射GLIS1-siRNA的牛胚胎可以发育到16-细胞期，但这些胚胎很难发育到32-细胞期以上。ZGA后转录的PDHA1和HSPA8基因在GLIS1下调的胚胎中表达水平较低。有可能GLIS1下调的胚胎不能启动ZGA。我们的结果表明GLIS1是牛胚胎着床前发育的一个重要因素。
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.