Abstract
For somatic cell nuclear transfer cytoplasts from metaphase II, oocytes are exclusively used. However, it is evident that certain reprogramming activities are present in oocytes even at earlier stages of maturation. These activities are, however, only poorly characterised. The main reason for this is that even the intrinsic oocyte processes are insufficiently understood. The mammalian oocyte is a highly specialised cell that harbours many specific characteristics. One of these is its particularly large size when compared to somatic cells. As the oocyte enters the growth phase its volume, as well as the amount of material, increases considerably. Thus, it is clear that the oocyte must possess the machinery to accomplish this incredible material accumulation. When the growth phase is completed, the transcription ceases and the oocyte becomes transcriptionally inactive. In our study, we have used the model system of oocyte fusion (transcribing × non-transcribing germinal vesicle (GV) stage oocytes) as a substitute for a somatic cell nuclear transfer schemes where the somatic cell nucleus would be introduced into a cytoplast obtained from a GV stage oocyte. We wanted to determine if the fully grown GV stage oocyte could induce reprogramming of transcriptionally active transferred nucleus by suppressing this activity. In order to evaluate possible changes in transcriptional properties after nuclear transfer, we also investigated the mechanism of transcriptional silencing taking place when the oocyte reaches its full size as well as the fate of the components namely of the RNA polymerase II (Pol II) transcriptional and splicing machinery. Here, we show that while the Pol II is degraded in fully grown GV stage oocytes and the splicing proteins undergo significant rearrangement, these oocytes are unable to induce similar changes in transcriptionally active nuclei even after a prolonged culture interval.
Similar content being viewed by others
References
Aoki F, Worrad DM, Schultz RM (1997) Regulation of transcriptional activity during the first and second cell cycles in the preimplantation mouse embryo. Dev Biol 181:296–307
Borsuk E, Milik E (2005) Fully grown mouse oocyte contains transcription inhibiting activity which acts through histone deacetylation. Mol Reprod Dev 701:509–515
Bouniol-Baly C, Hamraoui L, Guibert J, Beaujean N, Szöllösi MS, Debey P (1999) Differential transcriptional activity associate with chromatin configuration in fully grown mouse germinal vesicle oocytes. Biol Reprod 60:580–587
Bui HT, Wakayama S, Kishigami S, Kim JH, Van Thuan N, Wakayama T (2008) The cytoplasm of mouse germinal vesicle stage oocytes can enhance somatic cell nuclear reprogramming. Development 135:3935–3945
Byrne JA, Simonsson S, Western PS, Gurdon JB (2003) Nuclei of adult mammalian somatic cells are directly reprogrammed to oct-4 stem cell gene expression by amphibian oocytes. Curr Biol 13:1206–1213
Cho EJ, Kobor MS, Kim M, Greenblatt J, Buratowski S (2001) Opposing effects of Ctk1 kinase and Fcp1 phosphatase at Ser2 of the RNA polymerase II C-terminal domain. Genes Dev 15:3319–3329
De La Fuente R (2006) Chromatin modifications in the germinal vesicle (GV) of mammalian oocytes. Dev Biol 292:1–12
De La Fuente R, Eppig JJ (2001) Transcriptional activity of the mouse oocyte genome: companion granulosa cells modulate transcription and chromatin remodelling. Dev Biol 229:224–236
De La Fuente R, Viveiros MM, Burns KH, Adashi EY, Matzuk MM, Eppig JJ (2004) Major chromatin remodelling in the germinal vesicle (GV) of mammalian oocytes is dispensable for global transcriptional silencing but required for centromeric heterochromatin function. Dev Biol 275:447–458
Debey P, Szöllösi MS, Szöllösi D, Vautier D, Girousse A, Besombes D (1993) Competent mouse oocytes isolated from antral follicles exhibit different chromatin organization and follow different maturation dynamics. Mol Reprod Dev 36:59–74
Doyle O, Corden JL, Murphy C, Gall JG (2002) The distribution of RNA polymerase II largest subunit (RPB1) in the Xenopus germinal vesicle. J Struct Biol 140:154–166
Fulka H (2007) Changes in global histone acetylation pattern in somatic cell nuclei after their transfer into oocytes at different stages of maturation. Mol Reprod Dev 75:556–564
Fulka H, Mrazek M, Fulka J Jr (2004) Nucleolar dysfunction may be associated with infertility in humans. Fertil Steril 82:486–487
Hahn S (2004) Structure and mechanism of the RNA polymerase II transcription machinery. Nat Struct Mol Biol 11:394–403
Handwerger KE, Gall JG (2006) Subnuclear organelles: new insights into form and function. Trends Cell Biol 16:19–26
Kishigami S, Mizutani E, Ohta H, Hikichi T, Thuan NV, Wakayma S, Bui H-T, Wakayama T (2006) Significant improvement of mouse cloning technique by treatment with trichostatin A after somatic nuclear transfer. Biochem Biophys Res Commun 340:183–189
Komarnitsky P, Cho EJ, Buratowski S (2000) Different phosphorylated forms of RNA polymerase II and associated mRNA processing factors during transcription. Genes Dev 14:2452–2460
Lamond AI, Spector DL (2003) Nuclear speckles: a model for nuclear organelles. Nat Rev Mol Cell Biol 4:605–612
Mehlmann LM (2005) Stops and starts in mammalian oocytes: recent advances in understanding the regulation of meiotic arrest and oocyte maturation. Reproduction 130:791–799
Meinhart A, Kamenski T, Hoeppner S, Baumli S, Cramer P (2005) A structural perspective of CTD function. Genes Dev 19:1401–1415
Miyara F, Migne C, Dumont-Hassan M, Le Meur A, Cohen-Bacrie P, Aubriot FX, Glissant A, Nathan C, Douard S, Stanovici A, Debey P (2003) Chromatin configuration and transcriptional control in human and mouse oocytes. Mol Reprod Dev 64:458–470
Mohammed AA, Karasiewicz J, Modliński JA (2008) Developmental potential of selectively enucleated immature mouse oocytes upon nuclear transfer. Mol Reprod Dev 75:1269–1280
Motlik J, Kubelka M (1990) Cell-cycle aspects of growth and maturation of mammalian oocytes. Mol Reprod Dev 27:366–375
Motlik J, Crozet N, Fulka J (1984) Meiotic competence in vitro of pig oocytes isolated from early antral follicles. J Reprod Fertil 72:323–328
Ogushi S, Palmieri C, Fulka H, Saitou M, Miyano T, Fulka J Jr (2008) The maternal nucleolus is essential for early embryonic development in mammals. Science 319:613–616
Parfenov VN, Davis DS, Pochukalina GN, Kostyuchek D, Murti KG (2000) Nuclear distribution of RNA polymerase II in human oocytes from antral follicles: dynamics relative to the transcriptional state and association with splicing factors. J Cell Biochem 77:654–665
Picton H, Briggs D, Gosden R (1998) The molecular basis of oocyte growth and development. Mol Cell Endocrinol 145:27–37
Sorensen RA, Wassarman PM (1976) Relationship between growth and meiotic maturation of the mouse oocyte. Dev Biol 50:531–536
Struhl K (1998) Histone acetylation and transcriptional regulatory mechanisms. Genes Dev 12:599–606
Sun F, Fang H, Li R, Gao T, Zheng J, Chen X, Ying W, Sheng HZ (2007) Nuclear reprogramming: the zygotic transcription program is established through an “erase-and-rebuild” strategy. Cell Res 17:117–134
Swiech L, Kisiel K, Czolowska R, Zientarski M, Borsuk E (2007) Accumulation and dynamics of proteins of the MCM family during mouse oogenesis and the first embryonic cell cycle. Int J Dev Biol 51:283–295
Truchet S, Chebrout M, Djediat C, Wietzerbin J, Debey P (2004) Presence of permanently activated signal transducers and activators of transcription in nuclear interchromatin granules of unstimulated mouse oocytes and preimplantation embryos. Biol Reprod 71:1330–1339
Wakayama T (2007) Production of cloned mice and ES cells from adult somatic cells by nuclear transfer: how to improve cloning efficiency? J Reprod Dev 53:13–26
Xie SQ, Martin S, Guillot PV, Bentley DL, Pombo A (2006) Slicing speckles are not reservoirs of RNA polymerase II, but contain an inactive form, phosphorylated on Serine2 residues of the C-terminal domain. Mol Biol Cell 17:1723–1733
Acknowledgment
This work was supported by grant MSMT 1M0021620803 from the Ministry of Education of the Czech Republic and GACR 523/09/1878 from the Czech Science Foundation. We also appreciate helpful comments from Pavel Hozak.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Fulka, H., Novakova, Z., Mosko, T. et al. The inability of fully grown germinal vesicle stage oocyte cytoplasm to transcriptionally silence transferred transcribing nuclei. Histochem Cell Biol 132, 457–468 (2009). https://doi.org/10.1007/s00418-009-0625-x
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00418-009-0625-x