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Identifying MicroRNA and mRNA Expression Profiles in Embryonic Stem Cells Derived from Parthenogenetic, Androgenetic and Fertilized Blastocysts
Authors:Xiang-Shun Cui  Xing-Hui Shen  Shao-Chen Sun  Sun-Wha Cho  Young-Tae Heo  Yong-Kook Kang  Teruhiko Wakayama
Institution:1. Department of Animal Science, Chungbuk National University, Gaeshin-dong, Cheongju, Chungbuk 361-763, South Korea
2. Department of Animal Science, Chungbuk National University, Gaeshin-dong, Cheongju, Chungbuk 361-763, South Korea;Department of Histology and Embryology, Harbin Medical University, Harbin 150081, China
3. Department of Animal Science, Chungbuk National University, Gaeshin-dong, Cheongju, Chungbuk 361-763, South Korea;College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
4. Laboratory of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806 South Korea
5. Center for Developmental Biology, RIKEN Kobe, Kobe 650-0047, Japan
Abstract:MicroRNAs (miRNAs) are a class of highly conserved small non-coding RNA molecules that play a pivotal role in several cellular functions. In this study, miRNA and messenger RNA (mRNA) profiles were examined by Illumina microarray in mouse embryonic stem cells (ESCs) derived from parthenogenetic, androgenetic, and fertilized blastocysts. The global analysis of miRNA-mRNA target pairs provided insight into the role of miRNAs in gene expression. Results showed that a total of 125 miRNAs and 2394 mRNAs were differentially expressed between androgenetic ESCs (aESCs) and fertilized ESCs (fESCs), a total of 42 miRNAs and 87 mRNAs were differentially expressed between parthenogenetic ESCs (pESCs) and fESCs, and a total of 99 miRNAs and 1788 mRNAs were differentially expressed between aESCs and pESCs. In addition, a total of 575, 5 and 376 miRNA-mRNA target pairs were observed in aESCs vs. fESCs, pESCs vs. fESCs, and aESCs vs. pESCs, respectively. Furthermore, 15 known imprinted genes and 16 putative uniparentally expressed miRNAs with high expression levels were confirmed by both microarray and real-time RT-PCR. Finally, transfection of miRNA inhibitors was performed to validate the regulatory relationship between putative maternally expressed miRNAs and target mRNAs. Inhibition of miR-880 increased the expression of Peg3, Dyrk1b, and Prrg2 mRNA, inhibition of miR-363 increased the expression of Nfat5 and Soat1 mRNA, and inhibition of miR-883b-5p increased Nfat5, Tacstd2, and Ppapdc1 mRNA. These results warrant a functional study to fully understand the underlying regulation of genomic imprinting in early embryo development.
Keywords:Embryonic stem cell  Parthenogenetic  Androgenetic  Fertilized  Microarray  miRNA-mRNA network
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