Comparing spatial expression dynamics of bovine blastocyst under three different procedures: in-vivo, in-vitro derived, and somatic cell nuclear transfer embryos.

Hiroaki Nagatomo, H. Akizawa, Ayari Sada, Yasunori Kishi, K. Yamanaka, T. Takuma, Keisuke Sasaki, N. Yamauchi, Y. Yanagawa, M. Nagano, T. Kono, Masashi Takahashi, M. Kawahara
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引用次数: 15

Abstract

There has been no work on spatiotemporal transcriptomic differences of blastocysts using in vivo- and in vitro-derived, and somatic cell nuclear transfer (SCNT) embryos. Here, we first compared the lineage-differentially transcriptomic profiles of in vivo- and in vitro-derived embryos by microarray analysis using divided into inner cell mass (ICM)-and trophectoderm (TE)-side samples, as well as those derived from SCNT in order to explore lineage-differentially expressed genes that are associated with preimplantation development in cattle. The transcriptomic profiles of the ICM-specific and TE-specific genes were similar between in vitro-derived embryos and in vivo-derived embryos, whereas SCNT embryos exhibited unusual lineage-differentially gene expression regulation at the blastocyst stage. The genes expressed in a spatiotemporal manner between developmentally normal in-vivo derived blastocysts and developmentally abnormal SCNT blastocysts might play critical roles for preimplantation development. Comparing spatial expression dynamics of bovine blastocyst under three different procedures revealed that CIITA was expressed in ICM-side samples of all the embryo types. CIITA is known as the master regulator of major histocompatibility complexes (MHC) class II genes that express in antigen-presenting cells but its biological function in preimplantation embryo is still unknown in mammals. Knockdown of CIITA expression in in vitro-derived embryos did not affect cleavage, but disrupted development of embryos into the blastocyst stage. These findings provide the novel transcriptomic information on blastocyst formation, raising the possibility that immune function-related gene directly plays important roles in bovine preimplantation development.
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比较三种不同程序下牛囊胚的空间表达动态:体内、体外衍生和体细胞核移植胚胎。
目前还没有关于体内和体外来源的囊胚和体细胞核移植(SCNT)胚胎的时空转录组学差异的研究。在这里,我们首先通过微阵列分析比较了体内和体外来源的胚胎的谱系差异转录组谱,并将其分为内细胞团(ICM)和滋养外胚层(TE)侧样本,以及来自SCNT的样本,以探索与牛着床前发育相关的谱系差异表达基因。icm特异性和te特异性基因的转录组谱在体外胚胎和体内胚胎中是相似的,而SCNT胚胎在囊胚期表现出不同寻常的谱系差异基因表达调控。发育正常的体内源性囊胚和发育异常的SCNT囊胚之间的时空表达基因可能在着床前发育中起关键作用。比较三种不同处理方式下牛囊胚的空间表达动态,发现CIITA在所有胚胎类型的icm侧样品中均有表达。CIITA被认为是主要组织相容性复合体(MHC) II类基因在抗原提呈细胞中表达的主要调控因子,但其在哺乳动物着床前胚胎中的生物学功能尚不清楚。在体外来源的胚胎中,敲低CIITA表达不影响卵裂,但破坏胚胎进入囊胚期的发育。这些发现为胚泡形成提供了新的转录组学信息,提出了免疫功能相关基因在牛着床前发育中直接起重要作用的可能性。
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