Maternally inherited rRNA triggers de novo nucleolus formation in porcine embryos
| العنوان: | Maternally inherited rRNA triggers de novo nucleolus formation in porcine embryos |
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| المؤلفون: | Frantisek Strejcek, Olga Østrup, Katarina Jedlickova, Lazo Pendovski, Jozef Laurincik, Matej Murin, Martin Morovic, Michal Benc, Andrea Lucas-Hahn, Alexandra Bartkova |
| المصدر: | Zygote. 26:395-402 |
| بيانات النشر: | Cambridge University Press (CUP), 2018. |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | Male, 0301 basic medicine, Swine, Nucleolus, Fertilization in Vitro, Immunofluorescence, Silver stain, 03 medical and health sciences, Microscopy, Electron, Transmission, RNA Polymerase I, Transcription (biology), medicine, RNA polymerase I, Animals, In Situ Hybridization, Fluorescence, Genome, medicine.diagnostic_test, Reverse Transcriptase Polymerase Chain Reaction, Chemistry, Embryo, Cell Biology, Ribosomal RNA, Molecular biology, Blastocyst, 030104 developmental biology, RNA, Ribosomal, Autoradiography, Female, Maternal Inheritance, Cell Nucleolus, Developmental Biology, Fluorescence in situ hybridization |
| الوصف: | 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 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. |
| تدمد: | 1469-8730 0967-1994 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::259643a264e238176d6998e517e60df0Test https://doi.org/10.1017/s0967199418000400Test |
| حقوق: | CLOSED |
| رقم الانضمام: | edsair.doi.dedup.....259643a264e238176d6998e517e60df0 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14698730 09671994 |
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