Single Molecule RNA Localization and Translation in the Mammalian Oocyte and Embryo. Issue 19 (17th September 2021)

• Record Type: Journal Article
• Authors: Jansova, Denisa; Aleshkina, Daria; Jindrova, Anna; Iyyappan, Rajan; An, Qin; Fan, Guoping; Susor, Andrej
• Is Part Of: Journal of molecular biology. Volume 433:Issue 19(2021)
• Publication Date: 2021-09-17
• Subjects: mRNA -- translation -- imaging -- localization -- subcellular; NEBD nuclear envelope breakdown -- FISH fluorescence in situ hybridization -- PLA proximity ligation assay -- MS2-GFP a highly specific binding between the RNA stem–loop sequence and the coat protein of the MS2 bacteriophage with a green fluorescent protein -- FUNCAT fluorescent noncanonical amino acid tagging -- Puro-PLA puromycylation proximity ligation assay -- RNA-puro-PLA RNA fluorescence in situ hybridization and the puromycilation proximity ligation assay; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2021.167166
• Languages: English
• ISSNs: 0022-2836

Abstract

Graphical abstract: Highlights: Novel RNA-puro-PLA technique detects temporal and spatial translation of a single mRNA molecule in the subcellular context. A significant correlation trend in expression was identified using multiple quantitative methods. Translationally dormant and abundant Ccnb1 and Mos mRNAs clustering in the cytoplasm. Abstract: During oocyte growth the cell accumulates RNAs to contribute to oocyte and embryo development which progresses with ceased transcription. To investigate the subcellular distribution of specific RNAs and their translation we developed a technique revealing several instances of localized translation with distinctive regulatory implications. We analyzed the localization and expression of candidate non-coding and mRNAs in the mouse oocyte and embryo. Furthermore, we established simultaneous visualization of mRNA and in situ translation events validated with polysomal occupancy. We discovered that translationally dormant and abundant mRNAs CyclinB1 and Mos are localized in the cytoplasm of the fully grown GV oocyte forming cloud-like structures with consequent abundant translation at the center of the MII oocyte. Coupling detection of the localization of specific single mRNA molecules with their translation at the subcellular context is a valuable tool to quantitatively study temporal and spatial translation of specific target mRNAs to understand molecular processes in the developing cell.