Identification and rescue of a novel TUBB8 mutation that causes the first mitotic division defects and infertility
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EMBRYO BIOLOGY
Identification and rescue of a novel TUBB8 mutation that causes the first mitotic division defects and infertility Yanping Jia 1,2 & Kunming Li 1 & Caihong Zheng 3 & Yuanyuan Tang 1,4 & Dandan Bai 2 & Jiqing Yin 2 & Fengli Chi 1 & Yalin Zhang 2 & Yanhe Li 2 & Zhifen Tu 2 & Yu Wang 1 & Jiaping Pan 1 & Shanshan Liang 1 & Yi Guo 1 & Jingling Ruan 2 & Pengcheng Kong 1 & Bi Wu 1 & Ye Hu 1 & Hong Wang 2 & Wenqiang Liu 2 & Xiaoming Teng 1,2 & Shaorong Gao 2 Received: 5 May 2020 / Accepted: 8 September 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
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. Keywords TUBB8 . Mutation . Embryo development arrest . Infertility . Rescue
Yanping Jia, Kunming Li, Caihong Zheng and Yuanyuan Tang contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10815-020-01945-w) contains supplementary material, which is available to authorized users. * Wenqiang Liu [email protected]
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Clinical and Translational Research Center of Shanghai First Maternity and Infant Hospital, Shanghai Key Laboratory of Signaling and Disease Research, Frontier Sci
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