Faculty of Veterinary Medicine and Animal Science, Division of Reproduction, Department of Clinical Sciences, Swedish University of Agricultural Sciences, SLU, Uppsala, Sweden.
Mol Reprod Dev. 2012 Aug;79(8):525-40. doi: 10.1002/mrd.22058. Epub 2012 Jul 12.
Following insemination, spermatozoa are retained in the utero-tubal junction and isthmic region of the oviduct, where essential steps of capacitation are coordinated. Although a majority of the spermatozoa is exposed to similar conditions in the oviduct, the speed of the response varies depending on the individual male and the state of the spermatozoa. The present study evaluated individual boar variations in terms of the ability of spermatozoa to undergo tyrosine phosphorylation in response to isthmic oviductal fluid (ODF). Cryopreserved spermatozoa from four boars were incubated with pre- and post-ovulatory ODF for 6 hr. Sperm kinematics, global protein tyrosine phosphorylation, and dynamics of different phosphorylation patterns were analyzed at hourly interval. The percentage of phosphorylated spermatozoa in the pre-ovulatory ODF-treated group was significantly (P < 0.001) higher than in the other treatment groups. Motility, velocity, and protein tyrosine phosphorylation in spermatozoa in response to ODF and control media also showed differences between boars. Spermatozoa from all four boars showed strong expression of a 19-kDa phosphoprotein while spermatozoa from two boars showed additionally strong expression of a 32-kDa phosphoprotein when incubated with pre-ovulatory ODF. While phosphorylation of proteins in the acrosome and the equatorial segment of the sperm were noticed at an early stage during incubation with ODF, tail phosphorylation appeared at a later stage of capacitation. The results indicate individual variation between boars in terms of sperm proteins, including different phosphorylation patterns, in response to ODF, which might be related to fertility.
受精后,精子被保留在输卵管的子宫-输卵管交界处和峡部,在这里协调获能的基本步骤。虽然大多数精子暴露在输卵管中的相似条件下,但反应速度取决于个体的雄性和精子的状态。本研究评估了精子对峡部输卵管液(ODF)发生酪氨酸磷酸化的能力在个体公猪中的差异。从四只公猪中冷冻保存的精子与排卵前和排卵后 ODF 孵育 6 小时。在每小时的间隔分析精子运动学、总蛋白酪氨酸磷酸化以及不同磷酸化模式的动力学。在排卵前 ODF 处理组中,磷酸化精子的百分比显著(P<0.001)高于其他处理组。ODF 和对照培养基对精子的运动性、速度和蛋白酪氨酸磷酸化也显示出公猪之间的差异。当用排卵前 ODF 孵育时,所有四只公猪的精子都表现出强烈的 19-kDa 磷酸蛋白表达,而两只公猪的精子还表现出强烈的 32-kDa 磷酸蛋白表达。在用 ODF 孵育期间,在早期阶段就注意到顶体和精子赤道段的蛋白质磷酸化,而尾部磷酸化出现在获能的后期阶段。研究结果表明,在对 ODF 的反应中,公猪的精子蛋白存在个体间的差异,包括不同的磷酸化模式,这可能与生育能力有关。