title: RNA profiles of porcine embryos during genome activation reveal complex metabolic switch sensitive to in vitro conditions.
abstract: Fertilization is followed by complex changes in cytoplasmic composition and extensive chromatin reprogramming which results in the abundant activation of totipotent embryonic genome at embryonic genome activation (EGA). While chromatin reprogramming has been widely studied in several species, only a handful of reports characterize changing transcriptome profiles and resulting metabolic changes in cleavage stage embryos. The aims of the current study were to investigate RNA profiles of in vivo developed (ivv) and in vitro produced (ivt) porcine embryos before (2-cell stage) and after (late 4-cell stage) EGA and determine major metabolic changes that regulate totipotency. The period before EGA was dominated by transcripts responsible for cell cycle regulation, mitosis, RNA translation and processing (including ribosomal machinery), protein catabolism, and chromatin remodelling. Following EGA an increase in the abundance of transcripts involved in transcription, translation, DNA metabolism, histone and chromatin modification, as well as protein catabolism was detected. The further analysis of members of overlapping GO terms revealed that despite that comparable cellular processes are taking place before and after EGA (RNA splicing, protein catabolism), different metabolic pathways are involved. This strongly suggests that a complex metabolic switch accompanies EGA. In vitro conditions significantly altered RNA profiles before EGA, and the character of these changes indicates that they originate from oocyte and are imposed either before oocyte aspiration or during in vitro maturation. IVT embryos have altered content of apoptotic factors, cell cycle regulation factors and spindle components, and transcription factors, which all may contribute to reduced developmental competence of embryos produced in vitro. Overall, our data are in good accordance with previously published, genome-wide profiling data in other species. Moreover, comparison with mouse and human embryos showed striking overlap in functional annotation of transcripts during the EGA, suggesting conserved basic mechanisms regulating establishment of totipotency in mammalian development.
authors: Institute for Basic Medical Sciences, Faculty of Medicine, University of Oslo and Norwegian Center for Stem Cell Research, Oslo, Norway. osvarcova@gmail.com
authors: Olga
authors: O
authors: Østrup
authors: Gayla
authors: G
authors: Olbricht
authors: Esben
authors: E
authors: Østrup
authors: Poul
authors: P
authors: Hyttel
authors: Philippe
authors: P
authors: Collas
authors: Ryan
authors: R
authors: Cabot
mesh_terms: D000818:Animals; D046508:Culture Techniques; D004622:Embryo, Mammalian / Q000196:embryology* / Q000378:metabolism*; D005306:Fertilization / Q000235:genetics; D020869:Gene Expression Profiling; D016678:Genome / Q000235:genetics*; D006801:Humans; D007753:Laboratories; D051379:Mice; D012313:RNA / Q000235:genetics*; D012326:RNA Splicing; D015203:Reproducibility of Results; D017423:Sequence Analysis, RNA; D013045:Species Specificity; D013552:Swine
chemical_list: D012313:RNA
references: Cell Death Differ. 2010 Jun;17(6):931-41
references: 19960021
references: Genome Biol. 2003;4(5):P3
references: 12734009
references: Dev Biol. 2006 Nov 1;299(1):206-20
references: 16945363
references: J Neurosci. 2011 Mar 9;31(10):3914-25
references: 21389246
references: Mol Cell Biochem. 2010 Jul;340(1-2):91-6
references: 20174964
references: Mol Reprod Dev. 2010 Sep;77(9):812-9
references: 20722012
references: J Biol Chem. 2008 Mar 21;283(12):7877-84
references: 18187419
references: Apoptosis. 2012 Nov;17(11):1235-45
references: 22914926
references: J Cell Biol. 2012 Apr 16;197(2):179-87
references: 22508508
references: Biochem Biophys Res Commun. 2005 Dec 23;338(3):1359-67
references: 16263084
references: Nat Cell Biol. 2011 Sep 25;13(11):1368-75
references: 21947082
references: RNA. 2008 Jul;14(7):1318-27
references: 18515545
references: Cell Cycle. 2007 Jun 15;6(12):1510-20
references: 17581281
references: J Biol Chem. 2011 May 13;286(19):17091-102
references: 21385873
references: Dev Dyn. 2005 Sep;234(1):215-21
references: 16086359
references: Oncotarget. 2012 Feb;3(2):183-94
references: 22383394
references: Dev Cell. 2009 Mar;16(3):398-410
references: 19289085
references: Cell Mol Life Sci. 2013 Apr;70(8):1425-37
references: 22965566
references: RNA. 2010 Mar;16(3):516-28
references: 20089683
references: Acta Ophthalmol Scand. 2005 Aug;83(4):471-6
references: 16029273
references: PLoS One. 2011;6(9):e24444
references: 21909434
references: Reproduction. 2009 Jun;137(6):931-42
references: 19279200
references: Brief Funct Genomics. 2012 Jan;11(1):57-62
references: 22210853
references: Cytogenet Cell Genet. 1996;74(1-2):146-8
references: 8893823
references: Nat Cell Biol. 2008 Feb;10(2):160-9
references: 18193033
references: Zygote. 1997 May;5(2):117-25
references: 9276509
references: Mol Cell. 2007 Nov 30;28(4):624-37
references: 18042457
references: BMC Cancer. 2011 May 26;11:199
references: 21612665
references: Free Radic Biol Med. 2012 Mar 1;52(5):889-97
references: 22226830
references: Dev Dyn. 2004 Apr;229(4):733-44
references: 15042697
references: Dev Cell. 2004 Jan;6(1):117-31
references: 14723852
references: J Biol Chem. 1996 Dec 6;271(49):31106-14
references: 8940107
references: Methods Mol Biol. 2013;927:385-96
references: 22992930
references: PLoS Genet. 2011 Dec;7(12):e1002390
references: 22194695
references: J Anim Sci. 2004 Jan;82(1):102-8
references: 14753353
references: Proc Natl Acad Sci U S A. 2005 Dec 6;102(49):17582-7
references: 16314565
references: Curr Opin Cell Biol. 2002 Jun;14(3):313-8
references: 12067653
references: Mol Biol Rep. 2012 May;39(5):5135-41
references: 22167327
references: Biol Reprod. 2004 Oct;71(4):1230-43
references: 15175238
references: Biol Reprod. 2010 Nov;83(5):791-8
references: 20668257
references: Nucleic Acids Res. 2010 Sep;38(16):5581-93
references: 20421206
references: Mol Hum Reprod. 2005 Mar;11(3):183-8
references: 15709158
references: Development. 2011 Jun;138(11):2369-78
references: 21558383
references: Mol Cell Biol. 2011 May;31(10):2053-65
references: 21383061
references: Biol Reprod. 2000 Dec;63(6):1848-56
references: 11090457
references: J Assist Reprod Genet. 2012 Jan;29(1):3-10
references: 21989496
references: Mol Cell. 2012 Jul 13;47(1):76-86
references: 22793692
references: Fertil Steril. 2009 Sep;92(3):835-848
references: 19631936
references: Wiley Interdiscip Rev RNA. 2010 Sep-Oct;1(2):230-9
references: 21935887
references: Zygote. 2014 Aug;22(3):356-65
references: 23331714
references: J Biol Chem. 2002 Jul 12;277(28):25233-8
references: 11994298
references: Genes Dev. 2011 May 1;25(9):972-83
references: 21536736
references: Genome Res. 2008 Sep;18(9):1509-17
references: 18550803
references: Cell Death Differ. 2000 Apr;7(4):402-7
references: 10773825
references: J Anim Sci. 2011 Nov;89(11):3561-71
references: 21680790
references: PLoS Genet. 2011 Jul;7(7):e1002175
references: 21811414
references: Proc Natl Acad Sci U S A. 2006 Dec 12;103(50):18905-10
references: 17142320
references: J Biol Chem. 2003 Nov 28;278(48):47753-61
references: 14506283
references: Anim Reprod Sci. 2007 Jul;100(1-2):128-40
references: 16895747
references: Genome Res. 2011 Aug;21(8):1328-38
references: 21555364
references: Nature. 2012 Nov 15;491(7424):393-8
references: 23151582
references: Biol Reprod. 2005 Feb;72(2):373-83
references: 15469999
references: Cell Biochem Biophys. 2011 Jun;60(1-2):61-8
references: 21468693
references: Mol Neurodegener. 2011 Jul 30;6:56
references: 21801444
references: Mol Cell. 2012 Jun 8;46(5):650-61
references: 22560923
references: PLoS One. 2012;7(4):e35357
references: 22523586
references: Semin Cell Dev Biol. 2012 Jul;23(5):509-14
references: 22343089
references: Mol Cell. 2007 Aug 17;27(4):609-21
references: 17707232
references: Cold Spring Harb Perspect Biol. 2011 Sep 01;3(9):a002758
references: 21690213
references: Development. 2011 Sep;138(17):3699-709
references: 21775417
references: Development. 2009 Sep;136(18):3033-42
references: 19700615
references: J Biosci. 2006 Mar;31(1):137-55
references: 16595883
references: Reproduction. 2009 Jan;137(1):23-33
references: 18845624
references: Biol Reprod. 1998 May;58(5):1316-20
references: 9603270
references: Physiol Plant. 2012 May;145(1):5-17
references: 22023699
references: Dev Cell. 2010 Aug 17;19(2):220-31
references: 20708585
references: Nat Commun. 2011;2:227
references: 21407193
references: Neoplasia. 2010 Jan;12(1):61-8
references: 20072654
references: Nature. 2011 Mar 24;471(7339):473-9
references: 21179090
references: Cell Cycle. 2012 Mar 15;11(6):1183-94
references: 22391207
references: Arch Biochem Biophys. 2012 Jul 15;523(2):151-60
references: 22546510
references: PLoS One. 2012;7(7):e42138
references: 22848731
references: Science. 2003 May 9;300(5621):980-4
references: 12738865
references: Dev Dyn. 2009 Nov;238(11):2850-9
references: 19842174
references: Biol Reprod. 2002 Jan;66(1):112-9
references: 11751272
references: BMC Genomics. 2009 Jan 08;10:10
references: 19128516
references: Nat Genet. 2002 Dec;32(4):627-32
references: 12434153
references: Mol Cell Biol. 2003 Nov;23(21):7437-47
references: 14559993
references: Zygote. 2007 May;15(2):117-28
references: 17462104
references: Proteomics. 2010 Nov;10(22):4128-41
references: 21080498
references: J Biol Chem. 2000 Mar 31;275(13):9823-31
references: 10734137
references: PLoS One. 2012;7(9):e44861
references: 22984577
references: PLoS One. 2013;8(2):e56535
references: 23409192
references: Mol Cell Biol. 2003 Jul;23(14):4805-13
references: 12832468
references: Cytogenet Cell Genet. 1995;69(1-2):75-80
references: 7835093
references: Mol Cell Biol. 2007 Dec;27(24):8421-30
references: 17938197
references: Environ Mol Mutagen. 2009 Oct;50(8):666-71
references: 19593801
references: J Cell Biochem. 2012 Mar;113(3):808-14
references: 22021003
references: Reprod Biomed Online. 2007 Feb;14(2):175-83
references: 17298719
references: Biol Reprod. 2005 Jun;72(6):1437-51
references: 15703372