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Identification and rescue of a novel TUBB8 mutation that causes the first mitotic division defects and infertility

  • Embryo Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

Tubulin beta eight class VIII (TUBB8) is essential for oogenesis, fertilization, and pre-implantation embryo development in human. Although TUBB8 mutations were recently discovered in meiosis-arrested oocytes of infertile females, there is no effective therapy for this gene mutation caused infertility. Our study aims to further reveal the infertility-causing gene mutations in the patient’s family and to explore whether the infertility could be rescued by optimizing the conditions of embryo culture and finally achieve the purpose of making the patient pregnant.

Methods

Whole-exome sequence analysis and Sanger sequencing were performed on patients’ family members to screen and identify candidate mutant genes. Construction of plasmids, in vitro transcription, microinjection of disease-causing gene cRNA, and immunofluorescence staining were used to recapitulate the infertility phenotype observed in patients and to understand the pathogenic principles. Simultaneously, overexpression of mutant and wild-type cRNA of the candidate gene in mouse oocytes at either germinal vesicle (GV) or metaphase II (MII) stage was performed in the rescue experiment.

Results

We first identified a novel heritable TUBB8 mutation (c.1041C>A: p.N347K) in the coding region which specifically affects the first mitosis and causes the developmental arrest of early embryos in a three-generation family. We further demonstrated that TUBB8 mutation could lead to abnormal spindle assemble. And moreover, additional expression of wild-type TUBB8 cRNA in the mouse oocytes in which the mutant TUBB8 were expressed can successfully rescue the developmental defects of resulting embryo and produce full-term offspring.

Conclusions

Our study not only defines a novel mutation of TUBB8 causing the early cleavage arrest of embryos, but also provides an important basis for treating such female infertility in the future.

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Funding

This project was supported by the National Natural Science Foundation of China (81630035, 31721003, 31601177, 31501197, 31501183, 81671521, and 31430056), the Ministry of Science and Technology of China (Grants 2016YFA0100400, 2015CB964800, 2018YFC1003102, 2014CB964601, 2016YFC1000600, and 2015CB964503), the Shanghai Natural Science Foundation (Grant 16ZR1427200), the Western Medicine Guidance Project of Shanghai Science and Technology Commission (1741196), the Youth Innovation Promotion Association of Chinese Academy of Sciences [2020104 to C.Z.], the Shanghai Shenkang Clinical Innovation Three-year Action Plan Project (16CR3069B).

Author information

Author notes

  1. Yanping Jia, Kunming Li, Caihong Zheng and Yuanyuan Tang contributed equally to this work.

Authors and Affiliations

  1. Clinical and Translational Research Center of Shanghai First Maternity and Infant Hospital, School of Medicine, Tongji University, Shanghai, 200092, China

    Yanping Jia, Kunming Li, Yuanyuan Tang, Fengli Chi, Yu Wang, Jiaping Pan, Shanshan Liang, Yi Guo, Pengcheng Kong, Bi Wu, Ye Hu & Xiaoming Teng

  2. Clinical and Translational Research Center of Shanghai First Maternity and Infant Hospital, Shanghai Key Laboratory of Signaling and Disease Research, Frontier Science Center for Stem Cell Research, School of Life Sciences and Technology, Tongji University, Shanghai, 200092, China

    Yanping Jia, Dandan Bai, Jiqing Yin, Yalin Zhang, Yanhe Li, Zhifen Tu, Jingling Ruan, Hong Wang, Wenqiang Liu, Xiaoming Teng & Shaorong Gao

  3. Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, 100101, China

    Caihong Zheng

  4. The People’s Hospital of Rizhao City in Shandong Province, Rizhao, 276800, China

    Yuanyuan Tang

Authors
  1. Yanping Jia
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  2. Kunming Li
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  3. Caihong Zheng
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  4. Yuanyuan Tang
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  8. Yalin Zhang
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  9. Yanhe Li
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  10. Zhifen Tu
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  11. Yu Wang
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  12. Jiaping Pan
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  18. Ye Hu
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  19. Hong Wang
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  20. Wenqiang Liu
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  21. Xiaoming Teng
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  22. Shaorong Gao
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Corresponding authors

Correspondence to Wenqiang Liu, Xiaoming Teng or Shaorong Gao.

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The authors declare that they have no conflict of interest.

Ethical approval

This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of Shanghai First Maternity and Infant Hospital of Tongji University.

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All subjects participating in this study signed their informed consents.

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Jia, Y., Li, K., Zheng, C. et al. Identification and rescue of a novel TUBB8 mutation that causes the first mitotic division defects and infertility. J Assist Reprod Genet 37, 2713–2722 (2020). https://doi.org/10.1007/s10815-020-01945-w

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  • DOI: https://doi.org/10.1007/s10815-020-01945-w

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