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胱硫醚β-合酶通过确保正确的减数分裂纺锤体组装来保证卵母细胞的质量。

Cystathionine β-synthase is required for oocyte quality by ensuring proper meiotic spindle assembly.

机构信息

Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing, China.

Department of Pathology, Beijing Luhe Hospital, Capital Medical University, Beijing, China.

出版信息

Cell Prolif. 2022 Dec;55(12):e13322. doi: 10.1111/cpr.13322. Epub 2022 Aug 19.

Abstract

OBJECTIVES

Poor oocyte quality is detrimental to fertilization and embryo development, which causes infertility. Cystathionine β-synthase (CBS) is one of the key enzymes modulating the metabolism of homocysteine (Hcy). Studies have shown that CBS plays an important role in female reproduction. However, the role of CBS in regulating oocyte quality during meiotic maturation still needs further investigation.

MATERIALS AND METHODS

Immunohistochemistry, immunofluorescence, drug treatment, western blot, cRNA construct and in vitro transcription, microinjection of morpholino oligo and cRNA were performed for this study.

RESULTS

We found that CBS was expressed both in human and mouse oocytes of follicles. In mouse oocytes, CBS was distributed in the nucleus at germinal vesicle (GV) stage and localized to spindle from germinal vesicle breakdown (GVBD) to metaphase II (MII). The expression of CBS was reduced in ovaries and oocytes of aged mice. CBS depletion resulted in meiotic arrest, spindle abnormality and chromosome misalignment, disrupted kinetochore-microtubule attachments and provoked spindle assembly checkpoint (SAC). CBS was disassembled when microtubules were disrupted with nocodazole, and co-localized with the stabilized microtubules after taxol treatment. Furthermore, CBS depletion decreased the acetylation of α-tubulin.

CONCLUSIONS

These results reveal that CBS is required for the acetylation of α-tubulin to ensure proper spindle assembly in regulating oocyte quality during meiotic maturation.

摘要

目的

卵母细胞质量差不利于受精和胚胎发育,从而导致不孕。胱硫醚β-合酶(CBS)是调节同型半胱氨酸(Hcy)代谢的关键酶之一。研究表明 CBS 在女性生殖中发挥重要作用。然而,CBS 在调节减数分裂成熟过程中卵母细胞质量的作用仍需要进一步研究。

材料与方法

本研究采用免疫组化、免疫荧光、药物处理、western blot、cRNA 构建和体外转录、卵母细胞形态发生素寡核苷酸和 cRNA 的显微注射等方法。

结果

我们发现 CBS 在人卵泡卵母细胞和小鼠卵母细胞中均有表达。在小鼠卵母细胞中,CBS 在生发泡期(GV)呈核分布,从生发泡破裂(GVBD)到中期 II(MII)定位于纺锤体。衰老小鼠的卵巢和卵母细胞中 CBS 的表达减少。CBS 耗竭导致减数分裂停滞、纺锤体异常和染色体错位,破坏着丝粒-微管连接,并引发纺锤体组装检查点(SAC)。用诺考达唑破坏微管时,CBS 被解聚,用紫杉醇处理后与稳定的微管共定位。此外,CBS 耗竭降低了α-微管蛋白的乙酰化。

结论

这些结果表明,CBS 是调节减数分裂成熟过程中卵母细胞质量所必需的,以确保适当的纺锤体组装,需要α-微管蛋白的乙酰化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/22a7/9715357/03b907a7918c/CPR-55-e13322-g006.jpg

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