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Updated: Oct 2, 2026
Mouse Sperm Cryopreservation and Recovery using the I·Cryo Kit
Published on: December 12, 2011
Cryopreservation Disrupts TRIM21-Mediated AKAP3 Ubiquitination and Triggers Stress Granule Formation in Human Sperm
Yan Wang1, Huizhong Xu2, Wenhong Lu3
1Institute of Pediatric Research, Children's Hospital of Soochow University, Suzhou, Jiangsu, China.
Background:
Sperm cryopreservation causes cryodamage, but the roles of posttranslational modifications (PTMs) and stress granules (SGs) in this process remain unclear.
Objectives:
To investigate the molecular mechanism of cryodamage via the TRIM21-AKAP3 axis, PTMs, and SGs.
Materials And Methods:
Human sperm were divided into fresh and cryopreserved groups. Global ubiquitinome, phosphoproteome, and acetylome profiling were performed, combined with co-immunoprecipitation (Co-IP), Western blotting, and immunofluorescence for functional validation. Sperm cryopreservation with FAZ-3532 intervention was further conducted to verify SG-mediated injury.
Results:
Cryopreservation-induced SG formation in mature human sperm is reported here for the first time. TRIM21 acted as the E3 ubiquitin ligase for AKAP3, targeting K31/K425; cryopreservation redirected TRIM21 into SGs, reducing AKAP3 ubiquitination and causing its accumulation. AKAP3 showed differential phosphorylation (22 sites) and acetylation (K433). Inhibiting SG assembly with FAZ-3532 remodeled AKAP3 ubiquitination patterns and partially improved post-thaw progressive motility.
Discussion And Conclusion:
PTM networks and SGs are critical for sperm cryodamage. TRIM21, AKAP3, and G3BP1 are potential targets to optimize sperm cryopreservation protocols, advancing male fertility preservation.
