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Published on: December 28, 2021
Deletion of Ck2β gene causes germ cell development arrest and azoospermia in male mice
Qiu-Xia Liang1,2, Zhen-Bo Wang2,3, Wen-Long Lei2,3
1Department of Reproductive Medicine, Peking University Shenzhen Hospital, Shenzhen, Guangdong, China.
Objectives:
In humans, non-obstructive azoospermia (NOA) is a major cause of male infertility. However, the aetiology of NOA is largely unknown. Previous studies reported that protein CK2β was abundantly and broadly expressed in spermatogenic cells. Here, we investigate whether protein CK2β participates in spermatogenesis.
Materials And Methods:
In this study, we separated spermatogenic cells using STA-PUT velocity sedimentation, analysed the expression pattern of protein CK2β by immunoblotting, specifically deleted Ck2β gene in early-stage spermatogenic cells by crossing Ck2βfl mice with Stra8-Cre+ mice and validated the knockout efficiency by quantitative RT-PCR and immunoblotting. The phenotypes of Ck2βfl/Δ ;SCre+ mice were studied by immunohistochemistry and immunofluorescence. The molecular mechanisms of male germ cell development arrest were elucidated by immunoblotting and TUNEL assay.
Results:
Ablation of Ck2β gene triggered excessive germ cell apoptosis, germ cell development arrest, azoospermia and male infertility. Inactivation of Ck2β gene caused distinctly reduced expression of Ck2α' gene and CK2α' protein.
Conclusions:
Ck2β is a vital gene for germ cell survival and male fertility in mice.
Insights
The protein CK2β is essential for male fertility. Its absence in mice leads to germ cell death, developmental arrest, and infertility, highlighting its vital role in spermatogenesis.
Area of Science:
- Reproductive Biology
- Molecular Genetics
- Cell Biology
Background:
- Non-obstructive azoospermia (NOA) is a significant cause of male infertility with largely unknown etiology.
- Protein CK2β is known to be highly expressed in spermatogenic cells, suggesting a potential role in male reproduction.
Purpose of the Study:
- To investigate the role of protein CK2β in spermatogenesis.
- To determine if CK2β is involved in the development of non-obstructive azoospermia.
Main Methods:
- Spermatogenic cells were isolated using STA-PUT velocity sedimentation.
- CK2β gene deletion was performed in early-stage spermatogenic cells of mice (Ck2βfl crossed with Stra8-Cre+).
- Gene knockout efficiency was validated using quantitative RT-PCR and immunoblotting.
- Phenotypic analysis of knockout mice involved immunohistochemistry and immunofluorescence.
- Molecular mechanisms of germ cell development arrest were studied via immunoblotting and TUNEL assay.
Main Results:
- Ablation of the Ck2β gene resulted in excessive germ cell apoptosis and developmental arrest.
- Mice lacking functional Ck2β exhibited azoospermia and male infertility.
- Inactivation of Ck2β led to significantly reduced expression of the Ck2α' gene and its corresponding protein.
Conclusions:
- CK2β is a critical gene for the survival of germ cells in mice.
- CK2β plays a vital role in maintaining male fertility.
- The study identifies CK2β as a key factor in spermatogenesis and male reproductive health.
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