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Published on: October 13, 2018
Effects of mechanical stresses on sperm function and fertilization rate in mice
Xiao Shi1, Ting Wang1, Zhuo Lin Qiu1
1a Center for Reproductive Medicine, Department of Obstetrics and Gynecology , Nanfang Hospital, Southern Medical University , Guang Zhou , China.
Abstract:
In this study, we investigated whether any of the observed changes in mouse sperm function tests secondary to mechanical stresses (centrifugation and pipetting) correlate with sperm fertilization ability. Chinese Kunming mice were used as sperm and oocyte donors. Sperm samples were allocated evenly into centrifugation, pipette, and control groups. Sperm plasma membrane integrity (PMI), mitochondrial membrane permeability (MMP), baseline and stimulated intracellular ROS, and sperm fertilization ability were measured by hypo-osmotic swelling, flow cytometry, and fertilization tests. Parallel studies were conducted and all tests were repeated six times. Our results showed that after centrifugation, the progressive motility, average path velocity, and overall sperm motility and PMI decreased significantly (p < 0.05). In addition, the MMP level decreased significantly in viable sperm when the centrifugation condition reached 1,400 g × 15 minutes (p < 0.05). When pipetting was performed two or more times, progressive motility, average path velocity, and overall sperm motility decreased significantly (p < 0.05); when it was performed four or more times, sperm membrane integrity and intracellular basal ROS level of viable sperm was also significantly decreased (p < 0.05). In conclusion, various mechanical stresses seem to affect sperm function, however this does not appear to alter fertilization rate. Laboratory handling steps should be minimized to avoid unnecessary mechanical stresses being applied to sperm samples.
Insights
Mechanical stresses like centrifugation and pipetting impact mouse sperm function, reducing motility and membrane integrity. However, these stresses do not significantly affect the sperm
Area of Science:
- Reproductive Biology
- Sperm Physiology
- Andrology
Background:
- Mechanical stresses are common during sperm handling in laboratory settings.
- The impact of these stresses on sperm function and fertilization capacity requires further investigation.
Purpose of the Study:
- To determine if mechanical stresses (centrifugation, pipetting) affect mouse sperm function tests.
- To correlate changes in sperm function with sperm fertilization ability.
Main Methods:
- Sperm samples from Chinese Kunming mice were subjected to centrifugation and pipetting.
- Measurements included sperm plasma membrane integrity (PMI), mitochondrial membrane permeability (MMP), intracellular reactive oxygen species (ROS), and fertilization tests.
- Standardized laboratory procedures and repeated trials ensured data reliability.
Main Results:
- Centrifugation significantly decreased sperm motility and PMI.
- High-force centrifugation also reduced MMP in viable sperm.
- Repeated pipetting negatively affected sperm motility, PMI, and ROS levels.
- Despite functional impairments, fertilization rates remained unaffected.
Conclusions:
- Mechanical stresses significantly impair various sperm functions.
- These functional deficits do not translate to a reduced fertilization rate in vitro.
- Minimizing mechanical handling during sperm processing is recommended to preserve sample quality.
