Alkmin Diego V, Parrilla Inmaculada, Tarantini Tatiana, Parlapan Laura, Del Olmo David, Vazquez Juan M, Martinez Emilio A, Roca Jordi
Department of Medicine and Animal Surgery, University of Murcia, Murcia, Spain.
Department of Medicine and Animal Surgery, University of Murcia, Murcia, Spain.
Theriogenology. 2014 Aug;82(3):501-8. doi: 10.1016/j.theriogenology.2014.05.008. Epub 2014 May 20.
To improve the efficiency of porcine sperm sex sorting using flow cytometry, the aims of the present study were to determine the relevance of inter- and intraboar variability in sperm sortability and to evaluate the significance of ejaculate semen characteristics in such variability. In addition, the variability among boars in the ability of sex-sorted spermatozoa to survive liquid storage at 15 °C to 17 °C was also evaluated. In total, 132 ejaculates collected from 67 boars of different breeds that were housed at an artificial insemination center were used in three experiments. X- and Y-chromosome-bearing sperm were simultaneously separated according to the Beltsville sperm-sorting technology using a high-speed flow cytometer. In the first experiment, interboar variability in the ability of the ejaculated spermatozoa to undergo the flow-based sex-sorting procedure was observed; the ejaculates of nearly 15% of the boars (n = 67) did not exhibit well-defined X- and Y-chromosome-bearing spermatozoa peaks in the histogram, and the ejaculate sperm concentration demonstrated good predictive value for explaining this variation, as indicated by the area under the receiver operating characteristics curve (0.88, P < 0.001). In the second experiment, a certain degree of intraboar variability was observed only in the boars that showed poor sperm sortability (measured according to the presence or not a well-defined split together with sperm sortability parameters) in the first ejaculate (n = 3). In contrast, boars classified as having good sperm sortability in the first ejaculate (n = 5) maintained this condition in five ejaculates collected over the subsequent 5 months. In the third experiment, sex-sorted spermatozoa from boars with good sperm sortability (n = 5) remained viable and motile (above 70% in all boars) after 48 hours of storage at 15 °C to 17 °C, which may facilitate the commercial application of sex-sorted spermatozoa in swine artificial insemination programs.
为提高使用流式细胞术进行猪精子性别分选的效率，本研究的目的是确定公猪间和公猪内精子可分选性的差异相关性，并评估射精精液特征在这种差异中的重要性。此外，还评估了不同公猪的经性别分选的精子在15℃至17℃下液态保存时存活能力的差异。总共从一个人工授精中心饲养的67头不同品种的公猪采集了132份精液样本，用于三项实验。使用高速流式细胞仪，根据贝尔茨维尔精子分选技术同时分离出携带X和Y染色体的精子。在第一个实验中，观察到射精精子进行基于流式的性别分选程序的能力存在公猪间差异；近15%的公猪（n = 67）的射精样本在直方图中未显示出明确的携带X和Y染色体精子的峰值，射精精子浓度对解释这种差异具有良好的预测价值，受试者工作特征曲线下面积表明了这一点（0.88，P < 0.001）。在第二个实验中，仅在第一次射精时精子可分选性较差（根据是否存在明确的分离以及精子分选参数来衡量，n = 3）的公猪中观察到了一定程度的公猪内差异。相比之下，在第一次射精时被归类为精子可分选性良好的公猪（n = 5）在随后5个月内采集的5次射精样本中均保持了这种状态。在第三个实验中，精子可分选性良好的公猪（n = 5）的经性别分选的精子在15℃至17℃下保存48小时后仍保持活力和运动能力（所有公猪均高于70%），这可能有助于经性别分选的精子在猪人工授精程序中的商业应用。
