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Differential expression of microRNAs in 2-cell and 4-cell mouse embryos

  • Pei Wang (a1), Ji Cui (a1), Chun Zhao (a1), Lin Zhou (a1), Xirong Guo (a1), Rong Shen (a1), Junqiang Zhang (a2) and Xiufeng Ling (a2)...


In vitro fertilized (IVF) human embryos have a high incidence of developmental arrest before the blastocyst stage, therefore characterization of the molecular mechanisms that regulate embryo development is urgently required. Post-transcriptional control by microRNAs (miRNAs) is one of the most investigated RNA control mechanisms, and is hypothesized to be involved actively in developmental arrest in preimplantation embryos. In this study, we extracted total RNA from mouse 2-cell and 4-cell embryos. Using a miRNA microarray, 192 miRNAs were found to be differentially expressed in 4-cell embryos and 2-cell embryos; 122 miRNAs were upregulated and 70 were downregulated in 4-cell embryos. The microarray results were confirmed by real-time quantitative RT-PCR for six miRNAs (mmu-miR-467h, mmu-miR-466d-3p, mmu-miR-292–5p, mmu-miR-154, mmu-miR-2145, and mmu-miR-706). Cdca4 and Tcf12 were identified as miR-154 target genes by target prediction analysis. This study provides a developmental map for a large number of miRNAs in 2-cell and 4-cell embryos. The function of these miRNAs and the mechanisms by which they modulate embryonic developmental arrest require further study. The results of this study have potential applications in the field of reproductive medicine.


Corresponding author

All correspondence to: Xiufeng Ling or to Junqiang Zhang. State Key Laboratory of Reproductive Medicine, Department of Reproduction, Nanjing Maternity and Child Health Care Hospital Affiliated to Nanjing Medical University, Nanjing Medical University, Nanjing, Jiangsu Province, China. Tel:/Fax: +86 25 52226248. e-mail: or


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Abbott, A.L., Alvarez-Saavedra, E., Miska, E.A., Lau, N.C., Bartel, D.P., Horvitz, H.R. & Ambros, V. (2005). The let-7 microRNA family members mir-48, mir-84, and mir-241 function together to regulate developmental timing in Caenorhabditis elegans. Dev. Cell. 9, 403–14.
Abrams, E.W. & Mullins, M.C. (2009). zebrafish development: it's in the maternal genes. Curr. Opin. Genet. Dev. 19, 396403.
Alizadeh, Z., Kageyama, S. & Aoki, F. (2005). Degradation of maternal mRNA in mouse embryos: selective degradation of specific mRNAs after fertilization. Mol. Reprod. Dev. 72, 281–90.
Bartel, D.P. (2004). MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116, 281–97.
Bashirullah, A., Halsell, S.R., Cooperstock, R.L., Kloc, M., Karaiskakis, A., Fisher, W.W., Fu, W., Hamilton, J.K., Etkin, L.D. & Lipshitz, H.D. (1999). Joint action of two RNA degradation pathways controls the timing of maternal transcript elimination at the midblastula transition in Drosophila melanogaster. EMBO J. 18, 2610–20.
Bettegowda, A., Lee, K.B. & Smith, G.W. (2008). Cytoplasmic and nuclear determinants of the maternal-to-embryonic transition. Reprod. Fertil. Dev. 20, 4553.
Bushati, N. & Cohen, S.M. (2007). MicroRNA functions. Annu. Rev. Cell Dev. Biol. 23, 175205.
Bushati, N., Stark, A., Brennecke, J. & Cohen, S.M. (2008). Temporal reciprocity of miRNAs and their targets during the maternal-to-zygotic transition in Drosophila. Curr. Biol. 18, 501–6.
Cui, X.S., Li, X.Y., Jeong, Y.J., Jun, J.H. & Kim, N.H. (2006). Gene expression of cox5a, 5b, or 6b1 and their roles in preimplantation mouse embryos. Biol. Reprod. 74, 601–10.
De Renzis, S., Elemento, O., Tavazoie, S. & Wieschaus, E.F. (2007). Unmasking activation of the zygotic genome using chromosomal deletions in the Drosophila embryo. PLoS Biol. 5, e117.
Delić, D., Dkhil, M., Al-Quraishy, S. & Wunderlich, F. (2011). Hepatic miRNA expression reprogrammed by Plasmodium chabaudi malaria. Parasitol. Res. 108, 1111–21.
Farley, B.M. & Ryder, S.P. (2008). Regulation of maternal mRNAs in early development. Crit. Rev. Biochem. Mol. Biol. 43, 135–62.
Giraldez, A.J., Cinalli, R.M., Glasner, M.E., Enright, A.J., Thomson, J.M., Baskerville, S., Hammond, S.M., Bartel, D.P. & Schier, A.F. (2005). MicroRNAs regulate brain morphogenesis in zebrafish. Science 308, 833–8.
Giraldez, A.J., Mishima, Y., Rihel, J., Grocock, R.J., Van Dongen, S., Inoue, K., Enright, A.J. & Schier, A.F. (2006). Zebrafish miR-430 promotes deadenylation and clearance of maternal mRNAs. Science 312, 75–9.
Hamatani, T., Carter, M.G., Sharov, A.A. & Ko, M.S. (2004). Dynamics of global gene expression changes during mouse preimplantation development. Dev. Cell 6, 117–31.
Chen, Hong, Qian, Kun, Ping Tang, Zhou, Xing, Bianzhi, Chen, Haiping, Liu, Na, Huang, Xiaolin & Zhang, Suming. (2010). Bioinformatics and microarray analysis of microRNA expression profiles of murine embryonic stem cells, neural stem cells induced from ESCs and isolated from E8.5 mouse neural tube. Neurol. Res. 32, 603–13.
Houbaviy, H.B., Murray, M.F. & Sharp, P.A. (2003). Embryonic stem cell-specific microRNA. Dev. Cell 5, 351–8.
Zhang, Junqiang, Cui, Ji, Ling, Xiufeng, Xiuling, Li, Peng, Yuzhu, Xirong, Guo, Chin Heng, Boon & Tong, Guo Qing. (2009). Vitrification of mouse embryos at 2-cell, 4-cell and 8-cell stages by cryotop method. J. Assist. Reprod. Genet. 26 (11–12), 621–8.
Lawitts, J.A. & Biggers, J.D. (1991). Overcoming the 2-cell block by modifying standard components in a mouse embryo culture medium. Biol. Reprod. 45, 245–51.
Lee, R.C., Feinbaum, R.L. & Ambros, V. (1993). The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 75, 843–54.
Lian, H., Liu, W., Liu, Q., Jin, H., Sun, Y., Li, J., Xia, Z. & Gao, H. (2010). A laboratory-attenuated vesicular stomatitis virus induces apoptosis and alters the cellular microRNA expression profile in BHK cells. Arch. Virol. 155, 1643–53.
Lund, E., Liu, M., Hartley, R.S., Sheets, M.D. & Dahlberg, J.E. (2009). Deadenylation of maternal mRNAs mediated by miR-427 in Xenopus laevis embryos. RNA 15, 2351–63.
Redell, J.B., Liu, Y. & Dash, P.K. (2009). Traumatic brain injury alters expression of hippocampal microRNAs: potential regulators of multiple patho-physiological processes. J. Neurosci. Res. 87, 1435–48.
Reinhart, B.J., Slack, F.J., Basson, M., Pasquinelli, A.E., Bettinger, J.C., Rougvie, A.E., Horvitz, H.R. & Ruvkun, G. (2000). The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans. Nature 403, 901–6.
Rosa, A., Spagnoli, F.M. & Brivanlou, A.H. (2009). The miR-430/427/302 family controls mesendodermal fate specification via species-specific target selection. Dev. Cell 16, 517–27.
Shkumatava, A., Stark, A., Sive, H. & Bartel, D.P. (2009). Coherent but over lapping expression of microRNAs and their targets during vertebrate development. Genes Dev. 23, 466–81.
Su, N., Wang, Y., Qian, M. & Deng, M. (2010). Combinatorial regulation of transcription factors and microRNAs. BMC Syst. Biol. 4, 150.
Tadros, W. & Lipshitz, H.D. (2009). The maternal-to-zygotic transition: a play in two acts. Development 136, 3033–42.
Takada, S., Berezikov, E., Yamashita, Y., Lagos-Quintana, M., Kloosterman, W.P., Enomoto, M., Hatanaka, H., Fujiwara, S., Watanabe, H., Soda, M., Choi, Y.L., Plasterk, R.H., Cuppen, E. & Mano, H. (2006). Mouse microRNA profiles determined with a new and sensitive cloning method. Nucl. Acids Res. 34, e115.
Tang, F., Kaneda, M., O'Carroll, D., Hajkova, P., Barton, S.C., Sun, Y.A., Lee, C., Tarakhovsky, A., Lao, K. & Surani, M.A. (2007). Maternal microRNAs are essential for mouse zygotic development. Genes Dev. 21, 644–8.
Wang, L.L., Zhang, Z., Li, Q., Yang, R., Pei, X., Xu, Y., Wang, J., Zhou, S.F. & Li, Y. (2009). Ethanol exposure induces differential microRNA and target gene expression and teratogenic effects which can be suppressed by folic acid supplementation. Hum. Reprod. 24, 562–79.
Watanabe, T., Imai, H. & Minami, N. (2008). Identification and expression analysis of small RNAs during development. Methods Mol. Biol. 442, 173–85.
Wu, L., Fan, J. & Belasco, J.G. (2006). MicroRNAs direct rapid deadenylation of mRNA. Proc. Natl. Acad. Sci. USA 103, 4034–9.
Ling, Xiu Feng, Zhang, Jun Qiang, Cao, Shan Ren, Chen, Jie, Peng, Yuzhu, Guo, Xirong, Heng, Boon Chin, Tong, Guo Qing & Wang, Xinru. (2009). Effect of cryotop vitrification on preimplantation developmental competence of murine morula and blastocyst stage embryos. Reprod. BioMed. Online 19, 708–13.
Zeng, F., Baldwin, D.A. & Schultz, R.M. (2004). Transcript profiling during preimplantation mouse development. Dev. Biol. 272, 483–96.


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Differential expression of microRNAs in 2-cell and 4-cell mouse embryos

  • Pei Wang (a1), Ji Cui (a1), Chun Zhao (a1), Lin Zhou (a1), Xirong Guo (a1), Rong Shen (a1), Junqiang Zhang (a2) and Xiufeng Ling (a2)...


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