PMID- 30311594
OWN - NLM
STAT- MEDLINE
DCOM- 20190122
LR  - 20190122
IS  - 1469-8730 (Electronic)
IS  - 0967-1994 (Linking)
VI  - 26
IP  - 5
DP  - 2018 Oct
TI  - Maternally inherited rRNA triggers de novo nucleolus formation in porcine 
      embryos.
PG  - 395-402
LID - 10.1017/S0967199418000400 [doi]
AB  - SummaryThe 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 mug/ml for inhibition of RPI; 2.0 mug/ml for inhibition 
      of total transcription) and late 2-cell embryos were cultured to the late 4-cell 
      stage with 0.2 mug/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.
FAU - Morovic, Martin
AU  - Morovic M
AUID- ORCID: 0000-0001-6898-5692
AD  - 1Constantine the Philosopher University,Nitra,Slovak Republic.
FAU - Ostrup, Olga
AU  - Ostrup O
AD  - 2Centre for Genomic Medicine,Copenhagen University Hospital,Copenhagen,Denmark.
FAU - Strejcek, Frantisek
AU  - Strejcek F
AD  - 1Constantine the Philosopher University,Nitra,Slovak Republic.
FAU - Benc, Michal
AU  - Benc M
AD  - 1Constantine the Philosopher University,Nitra,Slovak Republic.
FAU - Murin, Matej
AU  - Murin M
AD  - 1Constantine the Philosopher University,Nitra,Slovak Republic.
FAU - Jedlickova, Katarina
AU  - Jedlickova K
AD  - 1Constantine the Philosopher University,Nitra,Slovak Republic.
FAU - Bartkova, Alexandra
AU  - Bartkova A
AD  - 1Constantine the Philosopher University,Nitra,Slovak Republic.
FAU - Lucas-Hahn, Andrea
AU  - Lucas-Hahn A
AD  - 5Institute of Farm Animal Genetics (FLI),Mariensee,Neustadt,Germany.
FAU - Pendovski, Lazo
AU  - Pendovski L
AD  - 6Ss. Cyril and Methodius University in Skopje,Republic of Macedonia.
FAU - Laurincik, Jozef
AU  - Laurincik J
AD  - 1Constantine the Philosopher University,Nitra,Slovak Republic.
LA  - eng
PT  - Journal Article
DEP - 20181012
PL  - England
TA  - Zygote
JT  - Zygote (Cambridge, England)
JID - 9309124
RN  - 0 (RNA, Ribosomal)
RN  - EC 2.7.7.6 (RNA Polymerase I)
SB  - IM
MH  - Animals
MH  - Autoradiography
MH  - Blastocyst/cytology/*physiology
MH  - Cell Nucleolus/*genetics/physiology
MH  - Female
MH  - Fertilization in Vitro
MH  - Genome
MH  - In Situ Hybridization, Fluorescence
MH  - Male
MH  - *Maternal Inheritance
MH  - Microscopy, Electron, Transmission
MH  - RNA Polymerase I/genetics/metabolism
MH  - RNA, Ribosomal/*genetics/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Swine
OTO - NOTNLM
OT  - Embryonic genome activation
OT  - Nucleolus
OT  - Porcine embryo
OT  - RNA polymerase I
OT  - rRNA
EDAT- 2018/10/13 06:00
MHDA- 2019/01/23 06:00
CRDT- 2018/10/13 06:00
PHST- 2018/10/13 06:00 [pubmed]
PHST- 2019/01/23 06:00 [medline]
PHST- 2018/10/13 06:00 [entrez]
AID - S0967199418000400 [pii]
AID - 10.1017/S0967199418000400 [doi]
PST - ppublish
SO  - Zygote. 2018 Oct;26(5):395-402. doi: 10.1017/S0967199418000400. Epub 2018 Oct 12.