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Published on: March 11, 2018
[Rho/ROCK signaling pathway and anti-cryodamage ability of human sperm]
Jia Tian1, Shao-Hua Zhang2, Ke Ma2
1Ningxia Human Sperm Bank / Key Laboratory for Fertility Preservation and Maintenance of the Ministry of Education, General Hospital of Ningxia Medical University, Yinchuan, Ningxia 750004, China.
Objective:
To investigate the influence of the Rho/ROCK signaling pathway on the anti-cryodamage ability of human sperm and provide some theoretical evidence for the development of high-efficiency semen cryoprotectants.
Methods:
We collected semen samples from 25 healthy males, each divided into a fresh, a normal cryopreservation control and an Rho-inhibition group. Before and after freezing, we detected sperm motility, viability, membrane integrity, morphology, DNA fragmentation index (DFI), acrosomal enzyme activity (AEA) and mitochondrial membrane potential (MMP) and determined the expressions of RhoA and ROCK proteins in the sperm by immunofluorescence staining.
Results:
Compared with the normal cryopreservation control, the frozen-thawed sperm of the Rho-inhibition group showed significantly increased sperm motility （ ［51.20 ± 7.70］% vs ［57.50 ± 6.83］%, P = 0.002), survival rate ( ［52.87 ± 5.07］% vs ［60.24 ± 5.53］%, P = 0.001), membrane integrity (［59.78±5.56］% vs ［67.10 ± 4.43］%, P = 0.001), percentage of morphologically normal sperm (［4.83 ± 1.11］% vs ［7.46 ± 1.28］, P = 0.001) and MMP (56.30 ± 4.28 vs 63.11 ± 2.97, P = 0.001), but decreased DFI (［27.64 ± 6.64］% vs ［18.87 ± 4.07］%, P = 0.001). There was no statistically significant difference in the AEA of the frozen-thawed sperm between the control and Rho-inhibition groups (97.65 ± 9.31 vs 98.30 ± 11.33, P > 0.05). Immunofluorescence staining revealed extensive expressions of RhoA and ROCK proteins in the head and neck of the sperm.
Conclusions:
The Rho/ROCK signaling pathway plays a role in the cryodamage to human sperm, and inhibiting the activity of Rho/ROCK can significantly improve the ability of sperm to resist cryodamage.
Insights
Inhibiting the Rho/ROCK pathway significantly enhances human sperm's ability to withstand cryopreservation damage. This finding offers a promising strategy for developing improved semen cryoprotectants.
Area of Science:
- Reproductive Biology
- Cell Signaling
- Cryobiology
Background:
- Cryopreservation of human sperm is crucial for fertility treatments.
- Sperm cryodamage affects motility, viability, and DNA integrity.
- The Rho/ROCK signaling pathway's role in sperm cryosurvival is not fully understood.
Purpose of the Study:
- To investigate the influence of the Rho/ROCK signaling pathway on human sperm's anti-cryodamage ability.
- To provide theoretical evidence for developing high-efficiency semen cryoprotectants.
Main Methods:
- Semen samples from 25 healthy males were analyzed.
- Samples were divided into fresh, cryopreservation control, and Rho-inhibition groups.
- Sperm parameters (motility, viability, integrity, morphology, DNA fragmentation, acrosomal enzyme activity, mitochondrial membrane potential) and RhoA/ROCK protein expression were assessed.
Main Results:
- The Rho-inhibition group showed significantly improved sperm motility, survival rate, membrane integrity, normal morphology, and mitochondrial membrane potential post-thaw.
- DNA fragmentation index (DFI) was significantly decreased in the Rho-inhibition group.
- No significant difference in acrosomal enzyme activity (AEA) was observed between groups.
Conclusions:
- The Rho/ROCK signaling pathway is implicated in human sperm cryodamage.
- Inhibiting Rho/ROCK activity enhances sperm cryoresistance.
- Targeting Rho/ROCK may lead to advanced cryoprotective strategies for human sperm.
