Waberski D, Magnus F, Ardón F, Petrunkina A M, Weitze K F, Töpfer-Petersen E
Institute for Reproductive Medicine, University of Veterinary Medicine Hannover, Foundation, Buenteweg 15, D-30559 Hannover, Germany.
Reproduction. 2006 Feb;131(2):311-8. doi: 10.1530/rep.1.00814.
In vitro short-term storage of boar semen for up to 72 h before insemination negatively affects fertility, but this often remains undetected during semen quality assessment. One important sperm function is the ability to form the functional sperm reservoir in the oviduct. In the present study, we used the modified oviductal explant assay to study sperm binding to oviductal epithelium in vitro in diluted boar semen stored for 24 or 72 h. First, we determined the kinetics of in vitro sperm binding to oviductal epithelium in relation to co-incubation time of sperm and oviductal tissue pieces. Then, we studied how the binding of sperm to oviductal epithelium was affected by in vitro semen storage and by differences among individual boars. Sperm binding after different incubation times was significantly higher when semen was stored 24 h than after 72-h storage (P < 0.05), and peaked at 30-90 min of incubation. Sperm binding differed between boars (n = 44), and was negatively correlated to the percentage of sperm with cytoplasmic droplets (R = -0.51, P < 0.001). There were no significant changes in motility, acrosome integrity and propidium iodide stainability during the 72-h storage period. However, sperm-binding indices were significantly lower after 72 h in vitro storage than after 24-h storage in sperm from boars with normal semen quality (P < 0.05); in contrast, the binding capacity of sperm from boars with higher percentages of morphologically altered sperm remained at a low level. The sperm-binding capacity of sperm from four of the five boars with known subfertility was lower than the mean binding index minus one standard deviation of the boar population studied here. It is concluded that changes in the plasma membrane associated with in vitro ageing reduce the ability of stored boar sperm to bind to the oviductal epithelium. This study shows the potential of sperm-oviduct binding as a tool to assess both male fertility and changes in sperm function associated with in vitro ageing.
在授精前对公猪精液进行长达72小时的体外短期保存会对生育能力产生负面影响,但这在精液质量评估过程中往往未被发现。精子的一项重要功能是在输卵管中形成功能性精子储存库的能力。在本研究中,我们使用改良的输卵管外植体试验,研究在体外保存24小时或72小时的稀释公猪精液中精子与输卵管上皮的结合情况。首先,我们确定了体外精子与输卵管上皮结合的动力学与精子和输卵管组织块共同孵育时间的关系。然后,我们研究了体外精液保存以及不同公猪个体差异如何影响精子与输卵管上皮的结合。当精液保存24小时时,不同孵育时间后的精子结合显著高于保存72小时后(P<0.05),且在孵育30 - 90分钟时达到峰值。不同公猪(n = 44)之间的精子结合存在差异,并且与有细胞质滴的精子百分比呈负相关(R = -0.51,P<0.001)。在72小时的保存期内,精子活力、顶体完整性和碘化丙啶染色性均无显著变化。然而,精液质量正常的公猪精子在体外保存72小时后的精子结合指数显著低于保存24小时后(P<0.05);相反,形态改变精子百分比更高的公猪精子的结合能力则维持在较低水平。五头已知生育力低下的公猪中有四头的精子结合能力低于此处研究的公猪群体平均结合指数减一个标准差。结论是,与体外老化相关的质膜变化降低了储存的公猪精子与输卵管上皮结合的能力。本研究表明精子 - 输卵管结合作为评估雄性生育力以及与体外老化相关的精子功能变化的工具的潜力。