A heterozygous ZP2 mutation causes zona pellucida defects and female infertility in mouse and human

Sai-Li Liu; Hai-Yang Zuo; Bing-Wang Zhao; Jia-Ni Guo; Wen-Bo Liu; Wen-Long Lei; Yuan-Yuan Li; Ying-Chun Ouyang; Yi Hou; Zhi-Ming Han; Wei-Zhou Wang; Qing-Yuan Sun; Zhen-Bo Wang

doi:10.1016/j.isci.2023.107828

A heterozygous ZP2 mutation causes zona pellucida defects and female infertility in mouse and human

iScience. 2023 Sep 4;26(10):107828. doi: 10.1016/j.isci.2023.107828. eCollection 2023 Oct 20.

Authors

Sai-Li Liu^{1

2}, Hai-Yang Zuo³, Bing-Wang Zhao^{1

2}, Jia-Ni Guo^{1

2}, Wen-Bo Liu¹, Wen-Long Lei¹, Yuan-Yuan Li¹, Ying-Chun Ouyang¹, Yi Hou¹, Zhi-Ming Han^{1

2

4

5}, Wei-Zhou Wang³, Qing-Yuan Sun⁶, Zhen-Bo Wang^{1

2

4

5}

Affiliations

¹ State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.
² University of Chinese Academy of Sciences, Beijing 101408, China.
³ The Six Medical Center of Chinese People's Liberation Army General Hospital, Beijing 100048, China.
⁴ Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China.
⁵ Beijing Institute for Stem Cell and Regenerative Medicine, Beijing 100101, China.
⁶ Guangzhou Key Laboratory of Metabolic Diseases and Reproductive Health, Guangdong-Hong Kong Metabolism & Reproduction Joint Laboratory, Reproductive Medicine Center, Guangdong Second Provincial General Hospital, Guangzhou 510317, China.

Abstract

The zona pellucida (ZP) is an extracellular glycoprotein matrix surrounding mammalian oocytes. Recently, numerous mutations in genes encoding ZP proteins have been shown to be possibly related to oocyte abnormality and female infertility; few reports have confirmed the functions of these mutations in living animal models. Here, we identified a novel heterozygous missense mutation (NM_001376231.1:c.1616C>T, p.Thr539Met) in ZP2 from a primary infertile female. We showed that the mutation reduced ZP2 expression and impeded ZP2 secretion in cell lines. Furthermore, we constructed the mouse model with the mutation (Zp2^T541M) using CRISPR-Cas9. Zp2^WT/T541M female mice had normal fertility though generated oocytes with the thin ZP, whereas Zp2^T541M female mice were completely infertile due to degeneration of oocytes without ZP. Additionally, ZP deletion impaired folliculogenesis and caused female infertility in Zp2^T541M mice. Our study not only expands the spectrum of ZP2 mutation sites but also, more importantly, increases the understanding of pathogenic mechanisms of ZP2 mutations.

Keywords: Genetics; Molecular biology; Physiology.