Hostname: page-component-76fb5796d-qxdb6 Total loading time: 0 Render date: 2024-04-25T09:03:09.748Z Has data issue: false hasContentIssue false
  • English
  • Français

Maternally inherited rRNA triggers de novo nucleolus formation in porcine embryos

Published online by Cambridge University Press:  12 October 2018

Martin Morovic Open the ORCID record for Martin Morovic [Opens in a new window] ,
Olga Østrup ,
Frantisek Strejcek ,
Michal Benc ,
Matej Murin ,
Katarina Jedlickova ,
Alexandra Bartkova ,
Andrea Lucas-Hahn ,
Lazo Pendovski  and
Jozef Laurincik
Martin Morovic*
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic
Olga Østrup
Affiliation:
Centre for Genomic Medicine, Copenhagen University Hospital, Copenhagen, Denmark
Frantisek Strejcek
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic
Michal Benc
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic Institute of Animal Science, Prague, Czech Republic
Matej Murin
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic The Czech Academy of Sciences, Institute of Animal Physiology and Genetics, Libechov, Czech Republic
Katarina Jedlickova
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic
Alexandra Bartkova
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic
Andrea Lucas-Hahn
Affiliation:
Institute of Farm Animal Genetics (FLI), Mariensee, Neustadt, Germany
Lazo Pendovski
Affiliation:
Ss. Cyril and Methodius University in Skopje, Republic of Macedonia
Jozef Laurincik
Affiliation:
Constantine the Philosopher University, Nitra, Slovak Republic The Czech Academy of Sciences, Institute of Animal Physiology and Genetics, Libechov, Czech Republic
*
Author for correspondence: Martin Morovic. Department of Zoology and Anthropology, Faculty of Natural Sciences, Constantine the Philosopher University in Nitra, Nabrezie mladeze 91, 949 74, Nitra, Slovakia. Tel: +421 37 6408 712. Fax: +421 37 6408 556. E-mail: mmorovic@ukf.sk
Get access Rights & Permissions [Opens in a new window]

Summary

The present study examines the role of RNA polymerase I (RPI)-mediated transcription, maternally inherited rRNA and nucleolar proteins in the resumption of fibrillogranular nucleoli during embryonic genome activation (EGA) in porcine embryos. Late 4-cell embryos were incubated in the absence (control) or presence of actinomycin D (AD) (0.2 μg/ml for inhibition of RPI; 2.0 μg/ml for inhibition of total transcription) and late 2-cell embryos were cultured to the late 4-cell stage with 0.2 μg/ml AD to block EGA. Embryos were then processed for reverse-transcriptase polymerase chain reaction (RT-PCR), and for autoradiography (ARG), transmission electron microscopy (TEM), fluorescence in situ hybridization (FISH), silver staining and immunofluorescence (for RPI). Embryos in the control group displayed extranucleolar and intranucleolar ARG labelling, and exhibited de novo synthesis of rRNA and reticulated functional nucleoli. Nucleolar proteins were located in large foci. After RPI inhibition, nucleolar precursors transformed into segregated fibrillogranular structures, however no fibrillar centres were observed. The localization of rDNA and clusters of rRNA were detected in 57.1% immunoprecipitated (IP) analyzed nucleoli and dispersed RPI; 30.5% of nuclei showed large deposits of nucleolar proteins. Embryos from the AD-2.0 group did not display any transcriptional activity. Nucleolar formation was completely blocked, however 39.4% of nuclei showed rRNA clusters; 85.7% of nuclei were co-localized with nucleolar proteins. Long-term transcriptional inhibition resulted in the lack of ARG and RPI labelling; 40% of analyzed nuclei displayed the accumulation of rRNA molecules into large foci. In conclusion, maternally inherited rRNA co-localized with rDNA and nucleolar proteins can initiate a partial nucleolar assembly, resulting in the formation of fibrilogranular structures independently on activation of RPI-mediated transcription.

Keywords

Embryonic genome activationNucleolusPorcine embryoRNA polymerase IrRNAPorcine embryo
Type
Research Article
Information
Zygote , Volume 26 , Issue 5 , October 2018 , pp. 395 - 402
Copyright
© Cambridge University Press 2018 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Angelier, N, Tramier, M, Louvet, E, Coppey-Moisan, M, Savino, TM, De Mey, JR Hernandez-Verdun, D (2005) Tracking the interactions of rRNA processing proteins during nucleolar assembly in living cells. Mol Biol Cell 16, 2862–2871.Google Scholar
Baran, V, Fléchon, JE Pivko, J (1996) Nucleologenesis in the cleaving bovine embryo: immunocytochemical aspects. Mol Reprod Dev 44, 63–70.Google Scholar
Baran, V, Brochard, V, Renard, JP Fléchon, JE (2001) Nopp 140 involvement in nucleologenesis of mouse preimplantation embryos. Mol Reprod Dev 59, 277–284.Google Scholar
Baran, V, Pavlok, A, Bjerregaard, B, Wrenzycki, C, Hermann, D, Philimonenko, VV, Lapathitis, G, Hozak, P, Niemann, H Motlik, J (2004) Immunolocalization of upstream binding factor and pocket protein p130 during final stages of bovine oocyte growth. Biol Reprod 70, 877–886.Google Scholar
Bjerregaard, B Maddox-Hyttel, P (2004) Regulation of ribosomal RNA gene expression in porcine oocytes. Anim Reprod Sci 82, 605–616.Google Scholar
Bjerregaard, B, Wrenzycki, Ch, Strejcek, F, Laurincik, J, Holm, P, Ochs, RL, Rosenkranz, Ch, Callesen, H, Rath, D, Niemann, H Maddox-Hyttel, P (2004) Expression of nucleolar-related proteins in porcine preimplantation embryos produced in vivo and in vitro . Biol Reprod 70, 867–876.Google Scholar
Bjerregaard, B, Pedersen, HG, Jakobsen, AS, Rickords, LF, Lai, L, Cheong, H.-T, Samuel, M, Prather, RS, Strejcek, F, Rasmussen, ZR, Laurincik, J, Niemann, H, Maddox-Hyttel, P Thomsen, PD (2007) Activation of ribosomal RNA genes in porcine embryos produced in vitro or by somatic cell nuclear transfer. Mol Reprod Dev 74, 35–41.Google Scholar
Dousset, T, Wang, C, Verheggen, C, Chen, D, Hern andez-Verdun, D Huang, S (2000) Initiation of nucleolar assembly is independent of RNA polymerase I transcription. Mol Biol Cell 11, 2705–2717.Google Scholar
Fair, T, Hyttel, P, Lonergan, P Boland, MP (2001) Immunolocalization of nucleolar proteins during bovine oocyte growth, meiotic maturation, and fertilization. Biol Reprod 64, 1516–1525.Google Scholar
Florman, HM Ducibella, T (2006). Fertilization in mammals. In Knobil and Neill’s Physiology of Reproduction , pp. 149–196. Elsevier, NY, USA.Google Scholar
Fulka, H Aoki, F (2016) Nucleolus precursor bodies and ribosome biogenesis in early mammalian embryos: old theories and new discoveries. Biol Reprod 94, 1–8.Google Scholar
Hannan, KM, Hannan, RD, Smith, SD, Jefferson, LS, Lun, M Rothblum, LI (2000a) Rb and p130 regulate RNA polymerase I transcription: Rb disrupts the interaction between UBF and SL-1. Oncogene 19, 4988.Google Scholar
Hannan, KM, Kennedy, BK, Cavanaugh, AH, Hannan, RD, Hirschler-Laszkiewicz, I, Jefferson, LS Rothblum, LI (2000b) RNA polymerase I transcription in confluent cells: Rb downregulates rDNA transcription during confluence-induced cell cycle arrest. Oncogene 19, 3487.Google Scholar
Hemberger, M, Dean, W Reik, W (2009) Epigenetic dynamics of stem cells and cell lineage commitment: digging Waddington’s canal. Nat Rev Mol Cell Biol 10, 526–557.Google Scholar
Hyttel, P Madsen, I (1987) Rapid method to prepare mammalian oocytes and embryos for transmission electron microscopy. Cells Tissues Organs 129, 12–14.Google Scholar
Hyttel, P, Laurincik, J, Rosenkranz, C, Rath, D, Niemann, H, Ochs, RL Schellander, K (2000) Nucleolar proteins and ultrastructure in preimplantation porcine embryos developed in vivo . Biol Reprod 63, 1848–1856.Google Scholar
Kiss, T (2001) Small nucleolar RNA‐guided post‐transcriptional modification of cellular RNAs. EMBO J 20, 3617–3622.Google Scholar
Kopecny, V, Landa, V Pavlok, A (1995) Localization of nucleic acids in the nucleoli of oocytes and early embryos of mouse and hamster: an autoradiographic study. Mol Reprod Dev 41, 449–458.Google Scholar
Kovalska, M, Hruska-Plochan, M, Østrup, O, Adamkov, M, Lehotsky, J, Strejcek, F, Statelova, D, Mikuskova, K, Varga, I Petrovicova, I (2012) The embryonic nucleologenesis during inhibition of major transcriptional activity in bovine preimplantation embryos. Biologia 67, 818–825.Google Scholar
Labrecque, R, Lodde, V, Dieci, C, Tessaro, I, Luciano, AM Sirard, MA (2015) Chromatin remodelling and histone mRNA accumulation in bovine germinal vesicle oocytes. Mol Reprod Dev 82, 450–462.Google Scholar
Laurincik, J, Thomsen, PD, Hay-Schmidt, A, Avery, B, Greve, T, Ochs, RL Hyttel, P (2000a) Nucleolar proteins and nuclear ultrastructure in preimplantation bovine embryos produced in vitro . Biol Reprod 62, 1024–1032.Google Scholar
Laurincik, J, Zakhartchenko, V, Avery, B, Stojkovic, M, Brem, G, Wolf, E, Muller, M Hyttel, P (2000b) Activation of ribosomal RNA genes in preimplantation in vitro‐produced and nuclear transfer bovine embryos. Reprod Domest Anim 35, 255–259.Google Scholar
Laurincik, J, Schmoll, F, Mahabir, E, Schneider, H, Stojkovic, M, Zakhartchenko, V, Prelle, K, Hendrixen, PJ, Voss, PL, Moeszlacher, GG, Avery, B, Dieleman, SJ, Besenfelder, U, Muller, M, Ochs, RL, Wolf, E, Schellander, K Maddox-Hyttel, P (2003) Nucleolar proteins and ultrastructure in bovine in vivo developed, in vitro produced, and parthenogenetic cleavage-stage embryos. Mol Reprod Dev 65, 73–85.Google Scholar
Lavrentyeva, E, Shishova, K, Kagarlitsky, G Zatsepina, O (2015) Localisation of RNAs and proteins in nucleolar precursor bodies of early mouse embryos. Reprod Fert Dev 29, 509–520.Google Scholar
Lawson, K, Larentowicz, L, Laury-Kleintop, L Gilmour, SK (2005) B23 is a downstream target of polyamine-modulated CK2. Mol Cell Biochem 274, 103–114.Google Scholar
Liebers, R, Rassoulzadegan, M Lyko, F (2014) Epigenetic regulation by heritable RNA. PLoS Genet 10, e1004296.Google Scholar
Lindner, LE (1993) Improvements in the silver-staining technique for nucleolar organizer regions (AgNOR). J Histochem Cytochem 41, 439–445.Google Scholar
Lykke-Andersen, K, Gilchrist, MJ, Grabarek, JB, Das, P, Miska, E Zernicka-Goetz, M (2008) Maternal Argonaute 2 is essential for early mouse development at the maternal-zygotic transition. Mol Biol Cell 19, 4383–4392.Google Scholar
Morimoto, H, Ozaki, A, Okamura, H, Yoshida, K, Amorim, BR, Tanaka, H, Kitamura, S Haneji, T (2007) Differential expression of protein phosphatase type 1 isotypes and nucleolin during cell cycle arrest. Cell Biochem Funct 25, 369–375.Google Scholar
Ogushi, S, Palmieri, C, Fulka, H, Saitou, M, Miyano, T Fulka, J (2008) The maternal nucleolus is essential for early embryonic development in mammals. Science 319, 613–616.Google Scholar
Østrup, O, Hall, V, Petkov, SG, Wolf, XA, Hyldig, S Hyttel, P (2009) From zygote to implantation: morphological and molecular dynamics during embryo development in the pig. Reprod Domest Anim 44, 39–49.Google Scholar
Petters, RM Wells, KD (1993) Culture of pig embryos. J Reprod Fert Sci 48, 61–73.Google Scholar
Reimer, G, Raska, I, Tan, EM Scheer, U (1987) Human autoantibodies: probes for nucleolus structure and function. Virchows Arch B 54, 131–143.Google Scholar
Shishova, KV, Lavrentyeva, EA, Dobrucki, JW Zatsepina, OV (2015) Nucleolus-like bodies of fully-grown mouse oocytes contain key nucleolar proteins but are impoverished for rRNA. Dev Biol 397, 267–281.Google Scholar
Suh, N Blelloch, R (2011) Small RNAs in early mammalian development: from gametes to gastrulation. Development 138, 1653–1661.Google Scholar
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, 271–277.Google Scholar
Svarcova, O, Laurincik, J, Avery, B, Mlyncek, M, Niemann, H Maddox‐Hyttel, P (2007) Nucleolar development and allocation of key nucleolar proteins require de novo transcription in bovine embryos. Mol Reprod Dev 74, 1428–1435.Google Scholar
Svarcova, O, Strejcek, F, Petrovicova, I, Avery, B, Pedersen, HG, Lucas‐Hahn, A, Niemann, H, Laurincik, J Maddox‐Hyttel, P (2008) The role of RNA polymerase I transcription and embryonic genome activation in nucleolar development in bovine preimplantation embryos. Mol Reprod Dev 75, 1095–1103.Google Scholar
Verheggen, C, Le Panse, S, Almouzni, G Hernandez-Verdun, D (1998) Presence of pre-rRNAs before activation of polymerase I transcription in the building process of nucleoli during early development of Xenopus laevis . J Cell Biol 142, 1167–1180.Google Scholar
Verheggen, C, Almouzni, G Hernandez-Verdun, D (2000) The ribosomal RNA processing machinery is recruited to the nucleolar domain before RNA polymerase I during Xenopus laevis development. J Cell Biol 149, 293–306.Google Scholar
Viuff, D, Hyttel, P, Avery, B, Vajta, G, Greve, T, Callesen, H Thomsen, PD (1998) Ribosomal ribonucleic acid is transcribed at the 4-cell stage in in vitro-produced bovine embryos. Biol Reprod 59, 626–631.Google Scholar
Viuff, D, Greve, T, Holm, P, Callesen, H, Hyttel, P Thomsen, PD (2002) Activation of the ribosomal RNA genes late in the third cell cycle of porcine embryos. Biol Reprod 66, 629–634.Google Scholar
Yerle, M, Lahbib-Mansais, Y, Pinton, P, Robic, A, Goureau, A, Milan, D Gellin, J (1997) The cytogenetic map of the domestic pig (Sus scrofa domestica). Mamm Genome 8, 592–607.Google Scholar
Zatsepina, O, Baly, C, Chebrout, M Debey, P (2003) The step-wise assembly of a functional nucleolus in preimplantation mouse embryos involves the cajal (coiled) body. Dev Biol 253, 66–83.Google Scholar