Nait Mouloud M, Ouennoughi F, Yaiche L, Kaidi R, Iguer-Ouada M
Département des Sciences Biologiques de l'Environnement, Faculté des Sciences de la Nature et de la Vie, Université de Bejaia, 06000 Bejaia, Algérie; Laboratoire Associé en Ecosystèmes Marins et Aquacoles, Faculté des Sciences de la Nature et de la Vie, Université de Bejaia, 06000, Bejaia, Algeria.
Département des Sciences Biologiques de l'Environnement, Faculté des Sciences de la Nature et de la Vie, Université de Bejaia, 06000 Bejaia, Algérie.
Theriogenology. 2017 Mar 15;91:44-54. doi: 10.1016/j.theriogenology.2016.12.020. Epub 2016 Dec 21.
The aim of this study was to assess the effects of female bovine plasma collected at different days of the reproductive cycle on epididymal spermatozoa motility and to test hypothesis that the subpopulations pattern of motile spermatozoa is affected by this treatment. Blood plasma samples were collected from five Holstein Friesian cows at different stages of the estrous cycle (days 0, 5, 10, 12 and 18), one pregnant cow and one adult bull and were diluted 1:9 (V/V) with normal saline. Female charcoal-treated plasma, Bull plasma and saline were used as controls. Semen samples were obtained from cauda epididymidis through retrograde flushing and diluted in saline to approximately 60 × 106 sperm/ml. The extended semen was diluted 1:2 (V/V) with tested media and motility was evaluated at 15 min and then every hour for 6 h using a computer-assisted semen analysis. Multivariate clustering procedure was applied to identify and quantify specific subpopulations within the semen samples. The statistical analysis clustered all the motile spermatozoa into three separate subpopulations with defined patterns of movement: Subpopulation 1 poorly motile and non-progressive spermatozoa (39.3%), subpopulation 2 including the fastest and the most vigorous spermatozoa (46.4%) and subpopulation 3 represented by slow, non-vigorous but linear spermatozoa (14.3%). Initially, sperm samples supplemented with female, male or female charcoal-treated plasma stimulated equally total motility and spermatozoa belonging to subpopulation 2 regardless of the estrous cycle stage. After 1-h incubation, the motility of these both categories of spermatozoa (total motile and those assigned to subpopulation 2) is enhanced and maintained more in day 12, 18 and pregnant cow plasma than in female plasma from earlier stage of the estrous cycle (day 0, 5 and 10), male plasma and female-charcoal treated plasma. In conclusion, the overall results showed that female plasma stimulated significantly sperm motility, especially at the late stage of the estrous cycle. Additionally, to the diverse compounds contained in blood plasma, progesterone may play a key role in such motility activation.
本研究的目的是评估在生殖周期不同天数采集的雌性牛血浆对附睾精子活力的影响，并检验活动精子亚群模式受该处理影响这一假设。从五头处于发情周期不同阶段（第0、5、10、12和18天）的荷斯坦弗里生奶牛、一头怀孕奶牛和一头成年公牛采集血浆样本，并用生理盐水按1:9（V/V）稀释。雌性经活性炭处理的血浆、公牛血浆和生理盐水用作对照。通过逆行冲洗从附睾尾获取精液样本，并用生理盐水稀释至约60×10⁶个精子/毫升。将稀释后的精液按1:2（V/V）用受试培养基稀释，并使用计算机辅助精液分析在15分钟时评估活力，然后每小时评估一次，共评估6小时。应用多变量聚类程序识别和量化精液样本中的特定亚群。统计分析将所有活动精子聚类为三个具有明确运动模式的独立亚群：亚群1为活动能力差且非进行性的精子（39.3%），亚群2包括最快和最活跃的精子（46.4%），亚群3以缓慢、不活跃但呈线性的精子为代表（14.3%）。最初，补充雌性、雄性或雌性经活性炭处理血浆的精子样本，无论发情周期阶段如何，均能同等程度地刺激总活力以及属于亚群2的精子。孵育1小时后，与发情周期早期（第0、5和10天）的雌性血浆、雄性血浆和雌性经活性炭处理血浆相比，这两类精子（总活动精子和分配到亚群2的精子）在第12、18天和怀孕奶牛血浆中的活力增强且维持时间更长。总之，总体结果表明，雌性血浆能显著刺激精子活力，尤其是在发情周期后期。此外，除了血浆中含有的多种化合物外，孕酮可能在这种活力激活中起关键作用。
