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5.8S 前 rRNA 成熟因子、有丝分裂期磷蛋白 6 是一种卵子质量和减数分裂所必需的雌性生育因子。

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

机构信息

Center for Reproductive Medicine, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.

State Key Lab of Reproductive Medicine, Nanjing Medical University, Nanjing, China.

出版信息

Cell Prolif. 2020 Mar;53(3):e12769. doi: 10.1111/cpr.12769. Epub 2020 Jan 31.

Abstract

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.

摘要

目的

M 期磷蛋白 6(MPP6)对于体细胞中 5.8S 前 rRNA 的成熟很重要,它被筛选为一种雌性生育因子。然而,MPP6 是否在卵母细胞减数分裂和生育中发挥作用尚不清楚。我们旨在解决这个问题。

材料和方法

使用带有核包膜(SN)或无核包膜(NSN)的小鼠卵母细胞进行所有实验。使用肽纳米颗粒介导的抗体转染来耗尽 MPP6。免疫荧光染色、免疫组织化学和活细胞追踪染色用于检查 MPP6 的定位,并在对照或 MPP6 耗尽后描述表型。高保真 PCR 和荧光原位杂交(FISH)用于检查 5.8S rRNAs 的定位和水平。Western blot 用于检查蛋白质水平。MPP6-EGFP mRNA 微注射用于进行挽救。

结果

MPP6 在卵巢和卵母细胞中丰富。MPP6 耗尽显著阻碍卵母细胞减数分裂。MPP6 耗尽增加了 5.8S 前 rRNA。衰老卵母细胞中的 MPP6 和 5.8S rRNA 的 mRNA 水平降低,MPP6 mRNA 注射部分增加了 5.8S rRNA 的成熟度并改善了卵母细胞的质量。

结论

MPP6 是小鼠卵母细胞中 5.8S rRNA 成熟、减数分裂和质量控制所必需的,MPP6 水平可能是卵母细胞质量的标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/41a1/7106954/4dfaa303a4af/CPR-53-e12769-g001.jpg

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