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Zygotic Genome Activation pp 63–81Cite as

Detection of miRNA in Mammalian Oocytes and Embryos

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1605))

Abstract

MicroRNAs (miRNAs) play a crucial role in the regulation of many post-transcriptional processes in reproductive cells. Regulation of maternal mRNA translation and activation of zygotic mRNA are essential to successful embryonic development. Moreover, the precise development of embryonic cell and/or tissue lineages requires temporal and spatial control of gene expression, mRNA abundance, and translation into proteins, which is in part regulated via miRNA. Here, we describe some key protocols that can be utilized to detect and quantify miRNA in in vitro produced oocytes and embryos.

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References

  1. Hale BJ, Keating AF, Yang CX, Ross JW (2015) Small RNAs: their possible roles in reproductive failure. In: Bronson R (ed) The male role in pregnancy loss and embryo implantation failure. Springer, Switzerland

    Google Scholar 

  2. Eichenlaub-Ritter U, Peschke M (2002) Expression in in vivo and in vitro growing and maturing oocytes: focus on regulation of expression at the translational level. Hum Reprod Update 8:21–41

    Article  CAS  PubMed  Google Scholar 

  3. Wrenzycki C, Herrmann D, Niemann H (2007) Messenger RNA in oocytes and embryos in relation to embryo viability. Theriogenology 68S:S77–S83

    Article  Google Scholar 

  4. Hossain M, Salilew-Wondim D, Schellander K, Tesfaye D (2012) The role of microRNAs in mammalian oocytes and embryos. Anim Reprod Sci 134:36–44

    Article  CAS  PubMed  Google Scholar 

  5. Hale BJ, Yang CX, Ross JW (2014) Small RNA regulation of reproductive function. Mol Reprod Dev 81:148–159

    Article  CAS  PubMed  Google Scholar 

  6. Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism and function. Cell 116:281–297

    Article  CAS  PubMed  Google Scholar 

  7. Brennecke J, Stark A, Russell RB, Cohen SM (2005) Principles of microRNA-target recognition. PLoS Biol 3(3):e85

    Article  PubMed  PubMed Central  Google Scholar 

  8. Doench JG, Sharp PA (2004) Specificity of microRNA target selection in translational repression. Genes Dev 18(5):504–511

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Bagga S, Bracht J, Hunter S, Massirer K, Holtz J, Eachus R, Pasquinelli AE (2005) Regulation by let-7 and lin-4 miRNAs results in target mRNA degradation. Cell 122(4):553–563

    Article  CAS  PubMed  Google Scholar 

  10. Giraldez AJ, Mishima Y, Rihel J, Grocock RJ, Van Dongen S, Inoue K, Enright AJ, Schier AF (2006) Zebrafish MiR-430 promotes deadenylation and clearance of maternal mRNAs. Science 312(5770):75–79

    Article  CAS  PubMed  Google Scholar 

  11. Tang F, Kaneda M, O’Carroll D, Hajkova P, Barton S, Sun YA, Lee C, Tarakhovsky A, Lao K, Surani MA (2007) Maternal microRNAs are essential for mouse zygotic development. Genes Dev 21:644–648

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Stark A, Brennecke J, Bushati N, Russell RB, Cohen SM (2005) Animal MicroRNAs confer robustness to gene expression and have a significant impact on 3′UTR evolution. Cell 123(6):1133–1146

    Article  CAS  PubMed  Google Scholar 

  13. Babak T, Zhang W, Morris Q, Blencowe BJ, Hughes TR (2004) Probing microRNAs with microarrays: tissue specificity and functional inference. RNA 10(11):1813–1819

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Sood P, Krek A, Zavolan M, Macino G, Rajewsky N (2006) Cell-type-specific signatures of microRNAs on target mRNA expression. Proc Natl Acad Sci USA 103(8):2746–2751

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Schier AF (2007) The maternal-zygotic transition: death and birth of RNAs. Science 316(5823):406–407

    Article  CAS  PubMed  Google Scholar 

  16. Rajewsky N (2006) microRNA target predictions in animals. Nat Genet 38(Suppl):S8–S13

    Article  CAS  PubMed  Google Scholar 

  17. Brevini TA, Cillo F, Antonini S, Tosetti V, Gandolfi F (2007) Temporal and spatial control of gene expression in early embryos of farm animals. Reprod Fertil Dev 19(1):35–42

    Article  CAS  PubMed  Google Scholar 

  18. Tadros W, Lipshitz HD (2005) Setting the stage for development: mRNA translation and stability during oocyte maturation and egg activation in Drosophila. Dev Dyn 232(3):593–608

    Article  CAS  PubMed  Google Scholar 

  19. Watanabe T, Takeda A, Mise K, Okuno T, Suzuki T, Minami N, Imai H (2005) Stage-specific expression of microRNAs during Xenopus development. FEBS Lett 579(2):318–324

    Article  CAS  PubMed  Google Scholar 

  20. Aboobaker AA, Tomancak P, Patel N, Rubin GM, Lai EC (2005) Drosophila microRNAs exhibit diverse spatial expression patterns during embryonic development. Proc Natl Acad Sci USA 102(50):18017–18022

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Biemar F, Zinzen R, Ronshaugen M, Sementchenko V, Manak JR, Levine MS (2005) Spatial regulation of microRNA gene expression in the Drosophila embryo. Proc Natl Acad Sci USA 102(44):15907–15911

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Wienholds E, Kloosterman WP, Miska E, Alvarez-Saavedra E, Berezikov E, de Bruijn E, Horvitz HR, Kauppinen S, Plasterk RH (2005) MicroRNA expression in zebrafish embryonic development. Science 309(5732):310–311

    Article  CAS  PubMed  Google Scholar 

  23. Bernstein E, Kim SY, Carmell MA, Murchison EP, Alcorn H, Li MZ, Mills AA, Elledge SJ, Anderson KV, Hannon GJ (2003) Dicer is essential for mouse development. Nat Genet 35(3):215–217

    Article  CAS  PubMed  Google Scholar 

  24. Murchison EP, Stein P, Xuan Z, Pan H, Zhang MQ, Schultz RM, Hannon GJ (2007) Critical roles for Dicer in the female germline. Genes Dev 21(6):682–693

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Ma J, Flemr M, Stein P, Berninger P, Malik R, Zavolan M, Svoboda P, Schultz RM (2010) MicroRNA activity is suppressed in mouse oocytes. Curr Biol 20(3):265–270

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Suh N, Baehner L, Moltzahn F, Melton C, Shenoy A, Chen J, Blelloch R (2010) MicroRNA function is globally suppressed in mouse oocytes and early embryos. Curr Biol 20(3):271–277

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Koshiol J, Wang E, Zhao Y, Marincola F, Landi MT (2010) Strengths and limitations of laboratory procedures for microRNA detection. Cancer Epidemiol Biomarkers Prev 19(4):907–911

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Wright EC, Hale BJ, Yang CX, Njoka JG, Ross JW (2016) MicroRNA-21 and PDCD4 expression during in vitro oocyte maturation in pigs. Reprod Biol Endocrinol 14:21

    Article  PubMed  PubMed Central  Google Scholar 

  29. Yang CX, Du ZQ, Wright EC, Rothschild MF, Prather RS, Ross JW (2012) Small RNA profile of the cumulus oocyte complex and early embryos in the pig. Biol Reprod 87(5):117

    Article  PubMed  Google Scholar 

  30. Yang CX, Wright EC, Ross JW (2012) Expression of RNA binding proteins DND1 and FXR1 in the porcine ovary and during oocyte maturation and early embryo development. Mol Reprod Dev 79:541–552

    Article  CAS  PubMed  Google Scholar 

  31. Li M, Xia Y, Gu Y, Zhang K, Lang Q, Chen L, Guan J, Luo Z, Chen H, Li Y, Li Q, Li X, Jiang A, Shuai S, Wang J, Zhu Q, Zhou X, Gao X, Li X (2010) MicroRNAome of porcine pre- and postnatal development. PLoS One 5:7

    Google Scholar 

  32. mirVanaTM miRNA Isolation Kit; Ambion. https://tools.thermofisher.com/content/sfs/manuals/cms_055423.pdf

  33. Rio D (2011) Northern blots for small RNAs and microRNAs. Adapted from RNA: a laboratory manual by Donald C Rio, Manuel Ares Jr, Gregory J Hannon, Timothy W Nilsen. CSHL Press, Cold Spring Harbor, NY

    Google Scholar 

  34. Reddy A, Zheng Y, Jagadeeswaran G, Macmil S, Graham W, Roe B, Desilva U, Zhang W, Sunkar R (2009) Cloning, characterization and expression analysis of porcine microRNAs. BMC Genomics 10:65

    Article  PubMed  PubMed Central  Google Scholar 

  35. TaqMan Gene Expression Cells-to-CT Kit; Ambion. https://tools.thermofisher.com/content/sfs/manuals/cms_056225.pdf

  36. TaqMan® Small RNA Assays; Applied Biosystems. https://tools.thermofisher.com/content/sfs/manuals/cms_042167.pdf

  37. Lin Y, Zeng Y, Zhang F, Xue L, Huang Z, Li W, Guo M (2013) Characterization of microRNA expression profiles and the discovery of novel microRNAs involved in cancer during human embryonic development. PLoS One 8:8

    Google Scholar 

  38. TaqMan MicroRNA Assays; Applied Biosystems. https://genome.med.harvard.edu/documents/qpcr/microRNATaqManAssayProtocol.pdf

  39. RT2 miRNA PCR Arrays; SABiosciences. http://www.sabiosciences.com/Manual/mirnapcrarray.pdf

  40. SOLiD Small RNA Expression Kit; Ambion. http://tools.thermofisher.com/content/sfs/manuals/cms_054973.pdf

  41. Deo M, Yu J, Chung K, Tippens M, Turner D (2006) Detection of mammalian microRNA expression by in situ hybridization with RNA oligonucleotides. Dev Dyn 235:2538–2548

    Article  CAS  PubMed  Google Scholar 

  42. Wright E (2012) MicroRNA expression and function during porcine oocyte maturation and early embryonic development. Dissertation, Iowa State University

    Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Animal Science, Iowa State University, 2356 Kildee Hall, Ames, IA, 50011, USA

    Malavika K. Adur, Benjamin J. Hale & Jason W. Ross

Authors
  1. Malavika K. Adur
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  2. Benjamin J. Hale
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  3. Jason W. Ross
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Corresponding author

Correspondence to Jason W. Ross .

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Editors and Affiliations

  1. Department of Animal and Poultry Sciences, Virginia Tech, Blacksburg, Virginia, USA

    Kiho Lee

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Adur, M.K., Hale, B.J., Ross, J.W. (2017). Detection of miRNA in Mammalian Oocytes and Embryos. In: Lee, K. (eds) Zygotic Genome Activation. Methods in Molecular Biology, vol 1605. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6988-3_5

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  • DOI: https://doi.org/10.1007/978-1-4939-6988-3_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6986-9

  • Online ISBN: 978-1-4939-6988-3

  • eBook Packages: Springer Protocols

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