Sperm Binding to Epithelial Oviduct Explants in Bulls with Different Nonreturn Rates Investigated with a New In Vitro Model1
Sperm Binding to Epithelial Oviduct Explants in Bulls with Different Nonreturn Rates Investigated with a New In Vitro Model1
复制标题
用新的体外模型研究不同不归率公牛的精子与上皮输卵管外植体的结合1
作者：
I. de Pauw;A. Van Soom;H. Laevens;S. Verberckmoes;A. de Kruif
建立了一种新的体外分析精子与输卵管上皮结合能力的方法，以确定这种结合能力是否可以用来预测不返回率(NRR)。精子结合率通过计数5，5′，6，6′-tetrachloro-1，1′，3，3′-tetraethylbenzimidazolyl-carbocyanine碘化物(JC-1)标记的附着于输卵管上皮的精子和通过图像分析程序测量输卵管外植体的表面积来评估。HEPES+酪蛋白乳酸丙酮酸(TALP)是一种比体外受精(IVF)-TALP、TICH-199+10%胎牛血清和TICM-199单独用来研究精子与输卵管外植体结合的更有用的培养液。表面积为20 000μm2的输卵管外植体比表面积为100 000μm2的输卵管外植体提供了更一致的结果。在精子膜完整性为60%的情况下，结合在0.1mm2输卵管上皮上的精子数与孵育24小时后各雄鼠的NRR呈正相关。精子与输卵管外植体结合的能力的测定可以成为一种可靠的体外方法来预测给定的父本的NRR。
Abstract A new in vitro method was developed for analyzing the capacity of sperm to bind to oviductal epithelium to determine whether this binding capacity could be used to predict nonreturn rates (NRR). Sperm binding was evaluated by counting 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolyl-carbocyanine iodide (JC-1)-labeled spermatozoa attached to oviductal epithelium and by measuring the surface area of the oviduct explants by means of an image analysis program. Hepes + Tyrode albumin lactate pyruvate (TALP) was a more useful medium than in vitro fertilization (IVF)-TALP, TCM-199 medium + 10% fetal calf serum, and TCM-199 medium alone for the investigation of sperm binding to oviductal explants. Oviduct explants with a surface area of ;lt20 000 μm2 provided more consistent results than did explants with a surface area of >100 000 μm2. A positive association was found between the loge transformed number of spermatozoa bound to 0.1 mm2 oviductal epithelium and the NRR of the respective sires after 24 h of coincubation, provided that the membrane integrity of the sperm sample was >60%. Determination of the capacity of sperm to bind to oviductal explants could become a reliable in vitro method for predicting the NRR of a given sire.
DOI：
10.1071/rd9950871
发表时间：
1995
期刊：
Reproduction, fertility, and development
影响因子：
--
作者：
P. Watson
通讯作者：
P. Watson
影响因子：
2.2
作者：
Ellington, JE;Samper, JC;Wright, RW
通讯作者：
Wright, RW