T-complex protein 1 subunit zeta-2 (CCT6B) deficiency induces murine teratospermia

Peiyin Yang; Wenjing Tang; Huiling Li; Rong Hua; Yan Yuan; Yue Zhang; Yunfei Zhu; Yiqiang Cui; Jiahao Sha

doi:10.7717/peerj.11545

T-complex protein 1 subunit zeta-2 (CCT6B) deficiency induces murine teratospermia

PeerJ. 2021 Jun 1:9:e11545. doi: 10.7717/peerj.11545. eCollection 2021.

Authors

Peiyin Yang^#¹, Wenjing Tang^#¹, Huiling Li¹, Rong Hua¹, Yan Yuan¹, Yue Zhang¹, Yunfei Zhu¹, Yiqiang Cui¹, Jiahao Sha¹

Affiliation

¹ Department of Histology and Embryology, Nanjing Medical University, Nanjing, Jiangsu, China.

^# Contributed equally.

Abstract

Background: The CCT complex is an important mediator of microtubule assembly and intracellular protein folding. Owing to its high expression in spermatids, CCT knockdown can disrupt spermatogenesis. In the present report, we therefore evaluated the in vivo functionality of the testis-specific CCT complex component CCT6B using a murine knockout model system.

Methods: A CRISPR/Cas9 approach was used to generate Cct6b^-/- mice, after which candidate gene expression in these animals was evaluated via qPCR and Western blotting. Testicular and epididymal phenotypes were assessed through histological and immunofluorescent staining assays, while a computer-assisted sperm analyzer was employed to assess semen quality.

Results: Cct6b^-/- mice were successfully generated, and exhibited no differences in development, fertility, appearance, testis weight, or sperm counts relative to control littermates. In addition, no differences in spermatogenesis were detected when comparingCct6b^+/+ and Cct6b^-/- testes. However, when progressive motility was analyzed, the ratio of normal sperm was significantly decreased in Cct6b^-/- male mice, with nuclear base bending being the primary detected abnormality. In addition, slight decreases in Cct4 and Cct7 expression were detected.

Conclusion: These data indicated that CCT6B is an important regulator of murine spermatogenesis, with the loss of this protein resulting in CCT complex dysfunction, providing a foundation for further studies.

Keywords: CCT6B; Gene knockout; Protein folding; Spermatogenesis.

Grants and funding

This work was supported by the National Key Research and Development Program of China (2018YFC1004201, 2018YFC1003500), National Natural Science Foundation of China (92068109). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.