Del Olmo E, Bisbal A, Maroto-Morales A, García-Alvarez O, Ramon M, Jimenez-Rabadan P, Martínez-Pastor F, Soler A J, Garde J J, Fernandez-Santos M R
SaBio IREC (CSIC - UCLM - JCCM), Campus Universitario s. n. 02071 Albacete, Spain.
Regional Center of Animal Selection and Reproduction (CERSYRA) JCCM, 13300 Valdepeñas, Spain.
Anim Reprod Sci. 2013 Apr;138(1-2):102-9. doi: 10.1016/j.anireprosci.2013.02.007. Epub 2013 Feb 22.
The present study aims to examine the predictive value of some sperm parameters on male fertility. Semen samples from six Manchega rams were collected and cryopreserved. Sperm quality was assessed after thawing and after 2h of incubation, either in the freezing extender (37°C) or after dilution in Synthetic Oviductal Fluid (SOF) (38°C, 5% CO2), attempting to mimic the physiological conditions of the female reproductive tract. The following sperm parameters were evaluated: motility and kinetic parameters by computer-assisted semen analyzer (CASA), and sperm viability (propidium iodide), mitochondrial membrane potential (JC-1), apoptotic-like membrane changes (YO-PRO-1), acrosomal status (PNA-FITC), and intracellular calcium (fluo-3) by flow cytometry. Results showed no significant differences between incubation media neither after thawing nor after incubation. There were no significant correlations between fertility and sperm parameters assessed by flow cytometry. However, after incubation in the freezing extender, sperm samples from males with poor fertility yielded less linearity and velocity (P<0.05) as indicated by motility parameters analyzed by CASA. These results indicate that kinematic sperm motility parameters evaluation by CASA might be useful to identify samples with poor fertility.
本研究旨在探讨某些精子参数对雄性生育能力的预测价值。采集了6只曼彻格公羊的精液样本并进行冷冻保存。在解冻后以及在冷冻稀释液(37°C)中孵育2小时后或在合成输卵管液(SOF)(38°C,5%二氧化碳)中稀释后孵育2小时后,评估精子质量,试图模拟雌性生殖道的生理条件。评估了以下精子参数:通过计算机辅助精液分析仪(CASA)评估活力和动力学参数,通过流式细胞术评估精子活力(碘化丙啶)、线粒体膜电位(JC-1)、凋亡样膜变化(YO-PRO-1)、顶体状态(PNA-FITC)和细胞内钙(fluo-3)。结果显示,孵育介质在解冻后和孵育后均无显著差异。通过流式细胞术评估的生育能力与精子参数之间没有显著相关性。然而,在冷冻稀释液中孵育后,生育能力差的雄性的精子样本的线性度和速度较低(P<0.05),这由CASA分析的活力参数表明。这些结果表明,通过CASA评估精子运动学活力参数可能有助于识别生育能力差的样本。