title: Effects of mechanical stresses on sperm function and fertilization rate in mice.
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.
authors.affiliation: a Center for Reproductive Medicine, Department of Obstetrics and Gynecology , Nanfang Hospital, Southern Medical University , Guang Zhou , China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Shi
authors.affiliation: a Center for Reproductive Medicine, Department of Obstetrics and Gynecology , Nanfang Hospital, Southern Medical University , Guang Zhou , China.
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.affiliation: a Center for Reproductive Medicine, Department of Obstetrics and Gynecology , Nanfang Hospital, Southern Medical University , Guang Zhou , China.
authors.forename: Zhuo Lin
authors.initials: ZL
authors.lastname: Qiu
authors.affiliation: b Center for Clinical Medicine Research, Nanfang Hospital , Southern Medical University , Guang Zhou , China.
authors.forename: Ke
authors.initials: K
authors.lastname: Li
authors.affiliation: a Center for Reproductive Medicine, Department of Obstetrics and Gynecology , Nanfang Hospital, Southern Medical University , Guang Zhou , China.
authors.forename: Liu
authors.initials: L
authors.lastname: Li
authors.affiliation: c Assisted Reproductive Technology Unit, Prince of Wales Hospital , The Chinese University of Hong Kong , Hong Kong.
authors.forename: Carol Pui Shan
authors.initials: CP
authors.lastname: Chan
authors.affiliation: a Center for Reproductive Medicine, Department of Obstetrics and Gynecology , Nanfang Hospital, Southern Medical University , Guang Zhou , China.
authors.forename: Si Mei
authors.initials: SM
authors.lastname: Chan
authors.affiliation: c Assisted Reproductive Technology Unit, Prince of Wales Hospital , The Chinese University of Hong Kong , Hong Kong.
authors.forename: Tian-Chiu
authors.initials: TC
authors.lastname: Li
authors.affiliation: a Center for Reproductive Medicine, Department of Obstetrics and Gynecology , Nanfang Hospital, Southern Medical University , Guang Zhou , China.
authors.forename: Song
authors.initials: S
authors.lastname: Quan
mesh_terms: D000818:Animals; D002462:Cell Membrane; D002463:Cell Membrane Permeability; D002498:Centrifugation; D005260:Female; D005306:Fertilization; D008297:Male; D051379:Mice; D051336:Mitochondrial Membranes / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D013048:Specimen Handling; D013081:Sperm Motility; D013094:Spermatozoa / Q000502:physiology*; D013314:Stress, Mechanical
chemical_list: D017382:Reactive Oxygen Species
keywords: Centrifugation; fertilization ability; pipetting; sperm function