RhoA improves cryopreservation of rooster sperm through the Rho/RhoA-associated kinase/cofilin pathway

文献类型： 外文期刊

第一作者： Heng, Nuo

作者： Heng, Nuo;Zhao, Zhi-Xian;Guo, Yong;Gao, Shan;Cai, De-Lin;Sheng, Xi-Hui;Wang, Xiang-Guo;Xing, Kai;Xiao, Long-Fei;Long, Cheng;Ni, He-Min;Qi, Xiao-Long;Heng, Nuo;Zhu, Hua-Bin;Fu, Bo-Fan

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关键词： RhoA; rooster sperm; semen quality; programmed freezing; ultrastructure; cryopreservation

期刊名称：POULTRY SCIENCE （ 影响因子：4.014； 五年影响因子：4.192 ）

ISSN： 0032-5791

年卷期： 2022 年 101 卷 10 期

页码：

收录情况： SCI

摘要： Cryopreservation of rooster sperm leads to relatively low semen quality due to cytoskeletal damage during the freeze-thawing process. This study aimed to explore how the addition of RhoA recombinant protein affected the viability and subcellular structure of rooster sperm after freeze-thawing and elucidated the molecular mechanisms of sperm cryopreservation. Semen quality and acrosome integrity testing revealed that the addition of 0.5 mg/mL RhoA recombinant protein to the cryoprotectant fluid significantly increased sperm motility, survival rate, linearity, straight-line velocity, and acrosome integrity after freeze-thawing (P < 0.05). Ultrastructure analysis of cryopreserved sperm showed structural damage to the sperm plasma membrane, nuclear membrane, and tail. However, compared to the control, these structural changes were reduced upon the addition of RhoA recombinant protein to the cryoprotective fluid (P < 0.05). Western blotting revealed that the expression of Rho/RhoA-associated kinase and p-cofilin was increased, and cofilin expression was decreased after sperm cryopreservation with recombinant RhoA protein. Treatment with Y-27632, a ROCK antagonist, suppressed ROCK and p-cofilin expression and decreased semen quality, acrosome integrity, and ultrastructure integrity. In summary, we have demonstrated a cryoprotective effect in spermatozoa involving the Rho/ROCK pathway during freeze-thawing. Furthermore, the addition of 0.5 mg/mL RhoA recombinant protein to the cryoprotective fluid improved rooster semen quality and subcellular structural homeostasis after freeze-thawing via the Rho/ ROCK pathway. This pathway may regulate the dynamic reorganization of the actin cytoskeleton by regulating the cofilin phosphorylation.

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