At the end of the growth phase, mouse antral follicle oocytes acquire full developmental competence. In the mouse, this event is marked by the transition from the so-called non-surrounded nucleolus (NSN) chromatin configuration into the transcriptionally quiescent surrounded nucleolus (SN) configuration, which is named after a prominent perinucleolar condensed chromatin ring. However, the SN chromatin configuration alone is not sufficient for determining the developmental competence of the SN oocyte. There are additional nuclear and cytoplamic factors involved, while a little is known about the changes occurring in the cytoplasm during the NSN/SN transition. Here, we report functional analysis of maternal ELAVL2 an AU-rich element binding protein. Elavl2 gene encodes an oocyte-specific protein isoform (denoted ELAVL2°), which acts as a translational repressor. ELAVL2° is abundant in fully grown NSN oocytes, is ablated during the NSN/SN transition and remains low during the oocyte-to-embryo transition (OET). ELAVL2° overexpression during meiotic maturation causes errors in chromosome segregation, indicating the significance of naturally reduced ELAVL2° levels in SN oocytes. On the other hand, during oocyte growth, prematurely reduced Elavl2 expression results in lower yields of fully grown and meiotically matured oocytes, suggesting that Elavl2 is necessary for proper oocyte maturation. Moreover, Elavl2 knockdown showed stimulating effects on translation in fully grown oocytes. We propose that ELAVL2 has an ambivalent role in oocytes: it functions as a pleiotropic translational repressor in efficient production of fully grown oocytes, while its disposal during the NSN/SN transition contributes to the acquisition of full developmental competence.

Institute of Animal Physiology and Genetics AS CR; Libechov Czech Republic

Institute of Molecular Genetics AS CR; Prague Czech Republic

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Edson MA, Nagaraja AK, Matzuk MM. The mammalian ovary from genesis to revelation. Endocr Rev. 2009;30:624–712. doi: 10.1210/er.2009-0012. PubMed DOI PMC

Tan JH, Wang HL, Sun XS, Liu Y, Sui HS, Zhang J. Chromatin configurations in the germinal vesicle of mammalian oocytes. Mol Hum Reprod. 2009;15:1–9. doi: 10.1093/molehr/gan069. PubMed DOI

De La Fuente R. Chromatin modifications in the germinal vesicle (GV) of mammalian oocytes. Dev Biol. 2006;292:1–12. doi: 10.1016/j.ydbio.2006.01.008. PubMed DOI

Mattson BA, Albertini DF. Oogenesis: chromatin and microtubule dynamics during meiotic prophase. Mol Reprod Dev. 1990;25:374–83. doi: 10.1002/mrd.1080250411. PubMed DOI

Inoue A, Nakajima R, Nagata M, Aoki F. Contribution of the oocyte nucleus and cytoplasm to the determination of meiotic and developmental competence in mice. Hum Reprod. 2008;23:1377–84. doi: 10.1093/humrep/den096. PubMed DOI

Zuccotti M, Ponce RH, Boiani M, Guizzardi S, Govoni P, Scandroglio R, Garagna S, Redi CA. The analysis of chromatin organisation allows selection of mouse antral oocytes competent for development to blastocyst. Zygote. 2002;10:73–8. doi: 10.1017/S0967199402002101. PubMed DOI

Gu L, Wang Q, Sun QY. Histone modifications during mammalian oocyte maturation: dynamics, regulation and functions. Cell Cycle. 2010;9:1942–50. doi: 10.4161/cc.9.10.11599. PubMed DOI

De La Fuente R, Viveiros MM, Burns KH, Adashi EY, Matzuk MM, Eppig JJ. Major chromatin remodeling in the germinal vesicle (GV) of mammalian oocytes is dispensable for global transcriptional silencing but required for centromeric heterochromatin function. Dev Biol. 2004;275:447–58. doi: 10.1016/j.ydbio.2004.08.028. PubMed DOI

Burns KH, Viveiros MM, Ren Y, Wang P, DeMayo FJ, Frail DE, Eppig JJ, Matzuk MM. Roles of NPM2 in chromatin and nucleolar organization in oocytes and embryos. Science. 2003;300:633–6. doi: 10.1126/science.1081813. PubMed DOI

Xia M, He H, Wang Y, Liu M, Zhou T, Lin M, Zhou Z, Huo R, Zhou Q, Sha J. PCBP1 is required for maintenance of the transcriptionally silent state in fully grown mouse oocytes. Cell Cycle. 2012;11:2833–42. doi: 10.4161/cc.21169. PubMed DOI

Zuccotti M, Merico V, Sacchi L, Bellone M, Brink TC, Stefanelli M, Redi CA, Bellazzi R, Adjaye J, Garagna S. Oct-4 regulates the expression of Stella and Foxj2 at the Nanog locus: implications for the developmental competence of mouse oocytes. Hum Reprod. 2009;24:2225–37. doi: 10.1093/humrep/dep191. PubMed DOI

Zuccotti M, Merico V, Sacchi L, Bellone M, Brink TC, Bellazzi R, Stefanelli M, Redi CA, Garagna S, Adjaye J. Maternal Oct-4 is a potential key regulator of the developmental competence of mouse oocytes. BMC Dev Biol. 2008;8:97. doi: 10.1186/1471-213X-8-97. PubMed DOI PMC

Ma JY, Li M, Luo YB, Song S, Tian D, Yang J, Zhang B, Hou Y, Schatten H, Liu Z, et al. Maternal factors required for oocyte developmental competence in mice: transcriptome analysis of non-surrounded nucleolus (NSN) and surrounded nucleolus (SN) oocytes. Cell Cycle. 2013;12:1928–38. doi: 10.4161/cc.24991. PubMed DOI PMC

Li L, Baibakov B, Dean J. A subcortical maternal complex essential for preimplantation mouse embryogenesis. Dev Cell. 2008;15:416–25. doi: 10.1016/j.devcel.2008.07.010. PubMed DOI PMC

Flemr M, Ma J, Schultz RM, Svoboda P. P-body loss is concomitant with formation of a messenger RNA storage domain in mouse oocytes. Biol Reprod. 2010;82:1008–17. doi: 10.1095/biolreprod.109.082057. PubMed DOI PMC

Monti M, Zanoni M, Calligaro A, Ko MS, Mauri P, Redi CA. Developmental arrest and mouse antral not-surrounded nucleolus oocytes. Biol Reprod. 2013;88:2. doi: 10.1095/biolreprod.112.103887. PubMed DOI PMC

Piqué M, López JM, Foissac S, Guigó R, Méndez R. A combinatorial code for CPE-mediated translational control. Cell. 2008;132:434–48. doi: 10.1016/j.cell.2007.12.038. PubMed DOI

Racki WJ, Richter JD. CPEB controls oocyte growth and follicle development in the mouse. Development. 2006;133:4527–37. doi: 10.1242/dev.02651. PubMed DOI

Kim JH, Richter JD. Measuring CPEB-mediated cytoplasmic polyadenylation-deadenylation in Xenopus laevis oocytes and egg extracts. Methods Enzymol. 2008;448:119–38. doi: 10.1016/S0076-6879(08)02607-4. PubMed DOI

Belloc E, Méndez R. A deadenylation negative feedback mechanism governs meiotic metaphase arrest. Nature. 2008;452:1017–21. doi: 10.1038/nature06809. PubMed DOI

Mendez R, Richter JD. Translational control by CPEB: a means to the end. Nat Rev Mol Cell Biol. 2001;2:521–9. doi: 10.1038/35080081. PubMed DOI

Richter JD. CPEB: a life in translation. Trends Biochem Sci. 2007;32:279–85. doi: 10.1016/j.tibs.2007.04.004. PubMed DOI

Mendez R, Barnard D, Richter JD. Differential mRNA translation and meiotic progression require Cdc2-mediated CPEB destruction. EMBO J. 2002;21:1833–44. doi: 10.1093/emboj/21.7.1833. PubMed DOI PMC

Bakheet T, Williams BR, Khabar KS. ARED 3.0: the large and diverse AU-rich transcriptome. Nucleic Acids Res. 2006;34:D111–4. doi: 10.1093/nar/gkj052. PubMed DOI PMC

Barreau C, Paillard L, Osborne HB. AU-rich elements and associated factors: are there unifying principles? Nucleic Acids Res. 2005;33:7138–50. doi: 10.1093/nar/gki1012. PubMed DOI PMC

Garneau NL, Wilusz J, Wilusz CJ. The highways and byways of mRNA decay. Nat Rev Mol Cell Biol. 2007;8:113–26. doi: 10.1038/nrm2104. PubMed DOI

Voeltz GK, Steitz JA. AUUUA sequences direct mRNA deadenylation uncoupled from decay during Xenopus early development. Mol Cell Biol. 1998;18:7537–45. PubMed PMC

Okano HJ, Darnell RB. A hierarchy of Hu RNA binding proteins in developing and adult neurons. J Neurosci. 1997;17:3024–37. PubMed PMC

Akamatsu W, Okano HJ, Osumi N, Inoue T, Nakamura S, Sakakibara S, Miura M, Matsuo N, Darnell RB, Okano H. Mammalian ELAV-like neuronal RNA-binding proteins HuB and HuC promote neuronal development in both the central and the peripheral nervous systems. Proc Natl Acad Sci U S A. 1999;96:9885–90. doi: 10.1073/pnas.96.17.9885. PubMed DOI PMC

Hinman MN, Lou H. Diverse molecular functions of Hu proteins. Cell Mol Life Sci. 2008;65:3168–81. doi: 10.1007/s00018-008-8252-6. PubMed DOI PMC

Sakai K, Kitagawa Y, Hirose G. Analysis of the RNA recognition motifs of human neuronal ELAV-like proteins in binding to a cytokine mRNA. Biochem Biophys Res Commun. 1999;256:263–8. doi: 10.1006/bbrc.1999.0282. PubMed DOI

Fan XC, Steitz JA. HNS, a nuclear-cytoplasmic shuttling sequence in HuR. Proc Natl Acad Sci U S A. 1998;95:15293–8. doi: 10.1073/pnas.95.26.15293. PubMed DOI PMC

Tenenbaum SA, Carson CC, Lager PJ, Keene JD. Identifying mRNA subsets in messenger ribonucleoprotein complexes by using cDNA arrays. Proc Natl Acad Sci U S A. 2000;97:14085–90. doi: 10.1073/pnas.97.26.14085. PubMed DOI PMC

Ince-Dunn G, Okano HJ, Jensen KB, Park WY, Zhong R, Ule J, Mele A, Fak JJ, Yang C, Zhang C, et al. Neuronal Elav-like (Hu) proteins regulate RNA splicing and abundance to control glutamate levels and neuronal excitability. Neuron. 2012;75:1067–80. doi: 10.1016/j.neuron.2012.07.009. PubMed DOI PMC

Good PJ. The role of elav-like genes, a conserved family encoding RNA-binding proteins, in growth and development. Semin Cell Dev Biol. 1997;8:577–84. doi: 10.1006/scdb.1997.0183. PubMed DOI

Wiszniak SE, Dredge BK, Jensen KB. HuB (elavl2) mRNA is restricted to the germ cells by post-transcriptional mechanisms including stabilisation of the message by DAZL. PLoS One. 2011;6:e20773. doi: 10.1371/journal.pone.0020773. PubMed DOI PMC

Calder MD, Madan P, Watson AJ. Bovine oocytes and early embryos express Staufen and ELAVL RNA-binding proteins. Zygote. 2008;16:161–8. doi: 10.1017/S096719940700456X. PubMed DOI

Calder MD, Watson PH, Watson AJ. Culture medium, gas atmosphere and MAPK inhibition affect regulation of RNA-binding protein targets during mouse preimplantation development. Reproduction. 2011;142:689–98. doi: 10.1530/REP-11-0082. PubMed DOI

Colegrove-Otero LJ, Devaux A, Standart N. The Xenopus ELAV protein ElrB represses Vg1 mRNA translation during oogenesis. Mol Cell Biol. 2005;25:9028–39. doi: 10.1128/MCB.25.20.9028-9039.2005. PubMed DOI PMC

Arthur PK, Claussen M, Koch S, Tarbashevich K, Jahn O, Pieler T. Participation of Xenopus Elr-type proteins in vegetal mRNA localization during oogenesis. J Biol Chem. 2009;284:19982–92. doi: 10.1074/jbc.M109.009928. PubMed DOI PMC

Stein P, Svoboda P, Schultz RM. Transgenic RNAi in mouse oocytes: a simple and fast approach to study gene function. Dev Biol. 2003;256:187–93. doi: 10.1016/S0012-1606(02)00122-7. PubMed DOI

Su AI, Cooke MP, Ching KA, Hakak Y, Walker JR, Wiltshire T, Orth AP, Vega RG, Sapinoso LM, Moqrich A, et al. Large-scale analysis of the human and mouse transcriptomes. Proc Natl Acad Sci U S A. 2002;99:4465–70. doi: 10.1073/pnas.012025199. PubMed DOI PMC

Levine TD, Gao F, King PH, Andrews LG, Keene JD. Hel-N1: an autoimmune RNA-binding protein with specificity for 3′ uridylate-rich untranslated regions of growth factor mRNAs. Mol Cell Biol. 1993;13:3494–504. PubMed PMC

Smallwood SA, Tomizawa S, Krueger F, Ruf N, Carli N, Segonds-Pichon A, Sato S, Hata K, Andrews SR, Kelsey G. Dynamic CpG island methylation landscape in oocytes and preimplantation embryos. Nat Genet. 2011;43:811–4. doi: 10.1038/ng.864. PubMed DOI PMC

Pillai RS, Artus CG, Filipowicz W. Tethering of human Ago proteins to mRNA mimics the miRNA-mediated repression of protein synthesis. RNA. 2004;10:1518–25. doi: 10.1261/rna.7131604. PubMed DOI PMC

Svoboda P. RNA silencing in mammalian oocytes and early embryos. Curr Top Microbiol Immunol. 2008;320:225–56. doi: 10.1007/978-3-540-75157-1_11. PubMed DOI

Barker A, Epis MR, Porter CJ, Hopkins BR, Wilce MC, Wilce JA, Giles KM, Leedman PJ. Sequence requirements for RNA binding by HuR and AUF1. J Biochem. 2012;151:423–37. doi: 10.1093/jb/mvs010. PubMed DOI

Pascale A, Govoni S. The complex world of post-transcriptional mechanisms: is their deregulation a common link for diseases? Focus on ELAV-like RNA-binding proteins. Cell Mol Life Sci. 2012;69:501–17. doi: 10.1007/s00018-011-0810-7. PubMed DOI PMC

Hinman MN, Zhou HL, Sharma A, Lou H. All three RNA recognition motifs and the hinge region of HuC play distinct roles in the regulation of alternative splicing. Nucleic Acids Res. 2013;41:5049–61. doi: 10.1093/nar/gkt166. PubMed DOI PMC

Fialcowitz-White EJ, Brewer BY, Ballin JD, Willis CD, Toth EA, Wilson GM. Specific protein domains mediate cooperative assembly of HuR oligomers on AU-rich mRNA-destabilizing sequences. J Biol Chem. 2007;282:20948–59. doi: 10.1074/jbc.M701751200. PubMed DOI PMC

Doller A, Schlepckow K, Schwalbe H, Pfeilschifter J, Eberhardt W. Tandem phosphorylation of serines 221 and 318 by protein kinase Cdelta coordinates mRNA binding and nucleocytoplasmic shuttling of HuR. Mol Cell Biol. 2010;30:1397–410. doi: 10.1128/MCB.01373-09. PubMed DOI PMC

Jain RG, Andrews LG, McGowan KM, Pekala PH, Keene JD. Ectopic expression of Hel-N1, an RNA-binding protein, increases glucose transporter (GLUT1) expression in 3T3-L1 adipocytes. Mol Cell Biol. 1997;17:954–62. PubMed PMC

Brennan CM, Steitz JA. HuR and mRNA stability. Cell Mol Life Sci. 2001;58:266–77. doi: 10.1007/PL00000854. PubMed DOI PMC

Simone LE, Keene JD. Mechanisms coordinating ELAV/Hu mRNA regulons. Curr Opin Genet Dev. 2013;23:35–43. doi: 10.1016/j.gde.2012.12.006. PubMed DOI PMC

Wang S, Kou Z, Jing Z, Zhang Y, Guo X, Dong M, Wilmut I, Gao S. Proteome of mouse oocytes at different developmental stages. Proc Natl Acad Sci U S A. 2010;107:17639–44. doi: 10.1073/pnas.1013185107. PubMed DOI PMC

Chen J, Melton C, Suh N, Oh JS, Horner K, Xie F, Sette C, Blelloch R, Conti M. Genome-wide analysis of translation reveals a critical role for deleted in azoospermia-like (Dazl) at the oocyte-to-zygote transition. Genes Dev. 2011;25:755–66. doi: 10.1101/gad.2028911. PubMed DOI PMC

Nejepinska J, Malik R, Filkowski J, Flemr M, Filipowicz W, Svoboda P. dsRNA expression in the mouse elicits RNAi in oocytes and low adenosine deamination in somatic cells. Nucleic Acids Res. 2012;40:399–413. doi: 10.1093/nar/gkr702. PubMed DOI PMC

Chalupníková K, Lattmann S, Selak N, Iwamoto F, Fujiki Y, Nagamine Y. Recruitment of the RNA helicase RHAU to stress granules via a unique RNA-binding domain. J Biol Chem. 2008;283:35186–98. doi: 10.1074/jbc.M804857200. PubMed DOI PMC

Flemr M, Svoboda P. Ribonucleoprotein localization in mouse oocytes. Methods. 2011;53:136–41. doi: 10.1016/j.ymeth.2010.08.005. PubMed DOI

Peritz T, Zeng F, Kannanayakal TJ, Kilk K, Eiríksdóttir E, Langel U, Eberwine J. Immunoprecipitation of mRNA-protein complexes. Nat Protoc. 2006;1:577–80. doi: 10.1038/nprot.2006.82. PubMed DOI

Vinopal S, Cernohorská M, Sulimenko V, Sulimenko T, Vosecká V, Flemr M, Dráberová E, Dráber P. γ-Tubulin 2 nucleates microtubules and is downregulated in mouse early embryogenesis. PLoS One. 2012;7:e29919. doi: 10.1371/journal.pone.0029919. PubMed DOI PMC

Poueymirou WT, Schultz RM. Differential effects of activators of cAMP-dependent protein kinase and protein kinase C on cleavage of one-cell mouse embryos and protein synthesis and phosphorylation in one- and two-cell embryos. Dev Biol. 1987;121:489–98. doi: 10.1016/0012-1606(87)90185-0. PubMed DOI

Hadjantonakis AK, Papaioannou VE. Dynamic in vivo imaging and cell tracking using a histone fluorescent protein fusion in mice. BMC Biotechnol. 2004;4:33. doi: 10.1186/1472-6750-4-33. PubMed DOI PMC

Functional canonical RNAi in mice expressing a truncated Dicer isoform and long dsRNA

The neglected part of early embryonic development: maternal protein degradation

Main constraints for RNAi induced by expressed long dsRNA in mouse cells