The zona pellucida (ZP) is an extracellular matrix universally surrounding mammalian eggs, which is essential for oogenesis, fertilization, and pre-implantation embryo development. Here, we identified two novel heritable mutations of ZP2 and ZP3, both occurring in an infertile female patient with ZP-abnormal eggs. Mouse models with the same mutations were generated by CRISPR/Cas9 gene editing system, and oocytes obtained from female mice with either single heterozygous mutation showed approximately half of the normal ZP thickness compared to wild-type oocytes. Importantly, oocytes with both heterozygous mutations showed a much thinner or even missing ZP that could not avoid polyspermy fertilization, following the patient’s pedigree. Further analysis confirmed that precursor proteins produced from either mutated ZP2 or ZP3 could not anchor to oocyte membranes. From these, we conclude that ZP mutations have dosage effects which can cause female infertility in humans. Finally, this patient was treated by intracytoplasmic sperm injection (ICSI) with an improved culture system and successfully delivered a healthy baby.
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We would like to thank the participant and families for their contribution to this study. We are grateful to our colleagues in the laboratory for their assistance with the experiments and in the preparation of this manuscript. We thank Dr. Jun Xiong for discussions about the in vitro experiments.
Conflict of interests
On behalf of all authors, the corresponding author states that there is no conflict of interests.
This project was supported by the National Natural Science Foundation of China (81630035, 31401247, 31501196, 31471392, 31601177, 31401266, 31501197, 31501183, 81671521, and 31430056), the Ministry of Science and Technology of China (Grants 2016YFA0100400, 2015CB964800, 2014CB964601, and 2015CB964503), Shanghai Natural Science Foundation (Grant 16ZR1427200), Shanghai Rising-Star Program (17QA1402700), the Municipal Education Commission of Shanghai (16CG17 and 16CG19) and the Program for Young Excellent Talents in Tongji University (Grants 2000219115, 2000219117, and 1515219023).
Wenqiang Liu, Kunming Li, Dandan Bai, and Jiqing Yin are contributed equally to this work.
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Liu, W., Li, K., Bai, D. et al. Dosage effects of ZP2 and ZP3 heterozygous mutations cause human infertility. Hum Genet 136, 975–985 (2017). https://doi.org/10.1007/s00439-017-1822-7