High-resolution microscopy of active ribosomal genes and key members of the rRNA processing machinery inside nucleolus-like bodies of fully-grown mouse oocytes

Shishova, K. V.; Khodarovich, Yu. M.; Lavrentyeva, Elena A.; Зацепина, О. В.

doi:10.1016/j.yexcr.2015.07.024

Cited by 15 publications

(16 citation statements)

References 49 publications

(98 reference statements)

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“…Nuclear exosome complex is required for the maturation of 5.8S rRNA in human somatic cells, while MPP6 is a nucleolus‐specific exosome co‐factor that binds preferentially to poly(C) and (U)‐rich ITS2 region to recruit the exosome . For SN FGOs that are transcriptionally quiescent, some researches suggested that rDNA‐containing nucleolus‐like bodies did not contain transcribed rRNA genes, pre‐rRNAs or pre‐ribosomes . Thus, we first wanted to verify whether pre‐rRNA existed within SN FGOs.…”

Section: Resultsmentioning

confidence: 99%

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The 5.8S pre‐rRNA maturation factor, M‐phase phosphoprotein 6, is a female fertility factor required for oocyte quality and meiosis

et al. 2020

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Objectives M‐phase phosphoprotein 6 (MPP6) is important for 5.8S pre‐rRNA maturation in somatic cells and was screened as a female fertility factor. However, whether MPP6 functions in oocyte meiosis and fertility is not yet known. We aimed to address this. Materials and Methods Mouse oocytes with surrounded nucleus (SN) or non‐surrounded nucleus (NSN) were used for all experiments. Peptide nanoparticle‐mediated antibody transfection was used to deplete MPP6. Immunofluorescence staining, immunohistochemistry and live tracker staining were used to examine MPP6 localization and characterize phenotypes after control or MPP6 depletion. High‐fidelity PCR and fluorescence in situ hybridization (FISH) were used to examine the localization and level of 5.8S rRNAs. Western blot was used to examine the protein level. MPP6‐EGFP mRNA microinjection was used to do the rescue. Results MPP6 was enriched within ovaries and oocytes. MPP6 depletion significantly impeded oocyte meiosis. MPP6 depletion increased 5.8S pre‐rRNA. The mRNA levels of MPP6 and 5.8S rRNA decreased within ageing oocytes, and MPP6 mRNA injection partially increased 5.8S rRNA maturation and improved oocyte quality. Conclusions MPP6 is required for 5.8S rRNA maturation, meiosis and quality control in mouse oocytes, and MPP6 level might be a marker for oocyte quality.

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Section: Resultsmentioning

confidence: 99%

“…[26][27][28] For SN FGOs that are transcriptionally quiescent, some researches suggested that rDNA-containing nucleolus-like bodies did not contain transcribed rRNA genes, pre-rRNAs or pre-ribosomes. [29][30][31] Thus, we first wanted to verify whether pre-rRNA existed within SN FGOs.…”

Section: The Female Fertility Factor Mpp6 Is Important For 58s Rrnmentioning

confidence: 99%

The 5.8S pre‐rRNA maturation factor, M‐phase phosphoprotein 6, is a female fertility factor required for oocyte quality and meiosis

et al. 2020

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“…It is retained as the so-called nucleolus-like body (NLB), sometimes referred to as the nucleolus precursor body (NPB). The NLB contains nucleolar proteins involved in all the main steps of ribosome biogenesis but not rRNA, presumably serving as a store of nucleolar proteins (Shishova et al 2015a(Shishova et al , 2015b. NLBs have been repeatedly documented to be essential for embryonic development (Ogushi et al 2008;Fulka and Aoki 2016); however, more recent data on their possible roles seem to be quite controversial.…”

Section: Nor Activity In Oocytes From Juvenile Birdsmentioning

confidence: 99%

Ribosomal RNA gene functioning in avian oogenesis

et al. 2016

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Despite long-term exploration into ribosomal RNA gene functioning during the oogenesis of various organisms, many intriguing problems remain unsolved. In this review, we describe nucleolus organizer region (NOR) activity in avian oocytes. Whereas oocytes from an adult avian ovary never reveal the formation of the nucleolus in the germinal vesicle (GV), an ovary from juvenile birds possesses both nucleolus-containing and non-nucleolus-containing oocytes. The evolutionary diversity of oocyte NOR functioning and the potential non-rRNA-related functions of the nucleolus in oocytes are also discussed.

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“…After the manipulation, the oocytes were washed in M2 medium, cultivated in CO 2 -incubator (5% CO 2 , 37°C) in the medium for in vitro maturation. The medium was prepared on the basis of DMEM C420 medium (PanEco, Russia) adding 15% fetal bovine serum (I31966-021; Gibco, USA), Gentamycin antibiotic (G1272; SigmaAldrich, USA) at 1.5 ME/ml concentration and PMSG1 ME/ml hormone (A036A02; Intervet, Germany) [5]. Observation of the development was started the next day 18 h after the operation.…”

mentioning

confidence: 99%

Determining the Range of Noninvasive Near-Infrared Femtosecond Laser Pulses for Mammalian Oocyte Nanosurgery

Osychenko¹,

Tochilo²,

Astafiev³

et al. 2017

Sovrem Tehnol Med

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The aim of the investigation was to determine a noninvasive range of near-infrared femtosecond laser pulse exposure for mouse oocyte intracellular nanosurgery and to estimate oocyte developmental competence in vitro depending on pulse energy and total exposure time.Materials and Methods. The model object was a preovulatory germinal vesicle mouse oocyte. Femtosecond laser radiation with a central wavelength of 780 nm was utilized in two modes: 1) single pulses with 30 fs duration and pulse energy up to 100 nJ; 2) trains of pulses with 100 fs duration, 80 MHz repetition rate (12.5 ns between pulses) and pulse energy from 0.5 to 2 nJ; total duration of the pulse trains ranged from 15 to 60 ms. Radiation was focused by a microscope objective lens (60×, 0.7 NA). Oocytes were cultured to the metaphase of the second meiotic division, which was used as invasiveness criteria. This stage was detected by polar bodies and metaphase plate formation.Results. The threshold of invasiveness was determined for a single femtosecond pulse exposure. Cytoplasmic membrane rupture and cell disruption occurred under the exposure of a single 30 fs pulse with 100 nJ energy. The cytoplasm and nucleus exposure to femtosecond pulse trains did not induce permanent damage to the oocytes. The rate of gas-vapor bubble formation was measured after the action of pulse trains on the cytoplasm and nucleus chromatin of mammalian oocytes. The pulse energy of 1 nJ is a threshold value for bubble formation in the cytoplasm, and 2 nJ in the nucleus. Exposure to laser pulse trains with 80 MHz repetition rate and 100 fs duration didn't change oocyte ability to achieve metaphase II stage within the studied range of pulse energies and pulse train durations.Conclusion. Surgery with femtosecond pulse trains at 80 MHz repetition rate and pulse energy not more than twice as high as the threshold of cavitation or vapor bubble formation does not affect oocyte development to the metaphase II stage. Femtosecond laser nanosurgery can be regarded as a minimally invasive method of intracellular surgery within the investigated range of pulse energies and exposure times.Key words: intracellular surgery; laser surgery; femtosecond laser; preovulatory oocytes.For contacts: Alina A. Osychenko, e-mail: alina.chemphys@gmail.com Laser Nanosurgery of Mammalian OocytesNanosurgical operations on the cells and embryos using femtosecond lasers with the wavelength in the near-infrared range is a challenging direction of modern biophotonics. The near-infrared range is in the region of biotissue transparency: it is not absorbed either by the water or organic substances of the cell (proteins, lipids, nucleic acids, carbohydrates) [1]. A high localization of the femtosecond pulse action in the area of laser beam waist (about 10 femtoliters volume) allows one to perform operations inside the cell/embryo on a separate organelle leaving intact the cell environment and causing no damage to the outer membrane [1][2][3]. Non-linear optical absorption of laser pulse energy by the cell material...

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High-resolution microscopy of active ribosomal genes and key members of the rRNA processing machinery inside nucleolus-like bodies of fully-grown mouse oocytes

Cited by 15 publications

References 49 publications

The 5.8S pre‐rRNA maturation factor, M‐phase phosphoprotein 6, is a female fertility factor required for oocyte quality and meiosis

The 5.8S pre‐rRNA maturation factor, M‐phase phosphoprotein 6, is a female fertility factor required for oocyte quality and meiosis

Ribosomal RNA gene functioning in avian oogenesis

Determining the Range of Noninvasive Near-Infrared Femtosecond Laser Pulses for Mammalian Oocyte Nanosurgery

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