MP70-10 IMPACTS OF COMMON SEMEN HANDLING METHODS ON SPERM FUNCTION (2016)

You have accessJournal of UrologyInfertility: Basic Research & Pathophysiology1 Apr 2016MP70-10 IMPACTS OF COMMON SEMEN HANDLING METHODS ON SPERM FUNCTION Alexander Travis, Cristina Cardona, Melissa Moody, Alana Simpson, and G. Charles Ostermeier Alexander TravisAlexander Travis More articles by this author , Cristina CardonaCristina Cardona More articles by this author , Melissa MoodyMelissa Moody More articles by this author , Alana SimpsonAlana Simpson More articles by this author , and G. Charles OstermeierG. Charles Ostermeier More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.1435AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES To become fertilization competent, sperm must undergo a maturational process known as capacitation. This process involves plasma membrane changes that occur in response to stimuli within the female tract. Methods currently used in semen analysis, and/or sperm preparation for ART could affect sperm membranes. Here, capacitation was evaluated using localization of the ganglioside GM1 (Cap-ScoreTMSperm Function Test). In particular we compared methods of liquefaction and washing used to reduce viscosity and determined their effects on capacitation. METHODS Three common methods to reduce viscosity were evaluated. Ejaculates were: 1) Incubated for 0.25, 1.25 or 2 hrs, 2) diluted 1:1 with Modified Human Tubal Fluid (Irvine Scientific; Santa Anna, CA) and then passed through a wide orifice transfer pipette (WOTP) or a Pasteur pipette (PP), 3) Enzymatically digested with chymotrypsin (chymo). Pilot studies revealed that passage through a hypodermic needle negatively affected motility and membrane integrity and was not studied further. After liquefaction, samples were washed and incubated under capacitating (CAP) and non-capacitating (NC) conditions. Cap-Score values were obtained via fluorescence microscopy. RESULTS Liquefaction time, dilution and pipetting did not alter Cap-Score. Control (incubation only), WOTP and PP treated samples had Cap-Scores of 41±4, 40±5, and 41±6 (n=5; CAP). A decrease in response to capacitating stimuli was observed when samples were liquefied using chymo (P=0.03). Control samples had Cap-Scores of 40±6 (n=5; CAP) whereas samples enzymatically liquefied had Cap-Scores of 31±4 (n=5; CAP). Because chymo is a protease that can cut membrane proteins we checked if the Cap-Score reduction resulted from an alteration in labeling. Samples not exposed to capacitation stimuli were compared and no difference was observed. Control and enzymatically liquefied samples had Cap-Scores of 22±4 and 21±5 (n=5; NC). These data support the view that treating semen with chymo can inhibit the ability of sperm to respond to capacitation stimuli. CONCLUSIONS Liquefaction times of up to 2 hours and mechanical liquefaction using WOTP and/or PP did not influence capacitation. In contrast, the use of enzymes such as chymo reduced the ability of sperm to capacitate, as measured by Cap-Score. These results demonstrate the importance of knowing how semen processing methods impact sperm function. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e909 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Alexander Travis More articles by this author Cristina Cardona More articles by this author Melissa Moody More articles by this author Alana Simpson More articles by this author G. Charles Ostermeier More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...