Ru360 protects against vitrification-induced oocyte meiotic defects by restoring mitochondrial function

Haowei Sun; Yaoyao Guo; Ruochun Yu; Jialun Wang; Youxue Liu; Hui Chen; Weijun Pang; Gongshe Yang; Guiyan Chu; Lei Gao

doi:10.1016/j.theriogenology.2023.04.004

Ru360 protects against vitrification-induced oocyte meiotic defects by restoring mitochondrial function

Theriogenology. 2023 Jul 1:204:40-49. doi: 10.1016/j.theriogenology.2023.04.004. Epub 2023 Apr 7.

Authors

Haowei Sun¹, Yaoyao Guo², Ruochun Yu³, Jialun Wang⁴, Youxue Liu⁵, Hui Chen⁶, Weijun Pang⁷, Gongshe Yang⁸, Guiyan Chu⁹, Lei Gao¹⁰

Affiliations

¹ Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, China. Electronic address: shwwj325@163.com.
² Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, China. Electronic address: GYY19981111@163.com.
³ Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, China. Electronic address: 2292212388@qq.com.
⁴ Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, China. Electronic address: 1067875173@qq.com.
⁵ Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, China. Electronic address: Lyx1867833@163.com.
⁶ Animal Husbandry Industry Test and Demonstration Center of Shaanxi Province, Jingyang, 713708, Shaanxi, China. Electronic address: chenhui089@163.com.
⁷ Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, China. Electronic address: pwj1226@nwafu.edu.cn.
⁸ Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, China. Electronic address: gsyang999@hotmail.com.
⁹ Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, China. Electronic address: guiyanchu@nwafu.edu.cn.
¹⁰ Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, China. Electronic address: gaolei5414@nwafu.edu.cn.

PMID: 37058855
DOI: 10.1016/j.theriogenology.2023.04.004

Abstract

Oocyte vitrification has been widely application in female fertility preservation. Recent studies found that vitrification of immature (germinal vesicle stage, GV) oocytes increased the risk of aneuploidy during meiotic maturation; however, the underlying mechanisms and the strategies to prevent this defect remain unexplored. In this study, we found that vitrification of GV oocytes decreased the first polarbody extrusion rate (90.51 ± 1.04% vs. 63.89 ± 1.39%, p < 0.05) and increased the aneuploid rate (2.50% vs. 20.00%, p < 0.05), accompanied with a series of defects during meiotic maturation, including aberrant spindle morphology, chromosome misalignment, incorrect Kinetochore-Microtubule attachments (KT-MTs) and weakened spindle assembly checkpoint protein complex (SAC) function. We also found that vitrification disrupted mitochondrial function by increasing mitochondrial Ca²⁺ levels. Importantly, inhibition of mitochondrial Ca²⁺ entry by 1 μM Ru360 significantly restored mitochondrial function and rescued the meiotic defects, indicating that the increase of mitochondrial Ca²⁺, at least, was a cause of meiotic defects in vitrified oocytes. These results shed light on the molecular mechanisms of oocyte vitrification-induced adverse effects of meiotic maturation and provided a potential strategy to improve oocyte cryopreservation protocols further.

Keywords: Aneuploidy; KT-MTs; Mitochondrial Ca(2+); Oocyte vitrification; Ru360; SAC.

MeSH terms

Aneuploidy
Animals
Cryopreservation / methods
Cryopreservation / veterinary
Female
Fertility Preservation* / veterinary
Mitochondria
Oocytes / physiology
Vitrification*