Mari G, Rizzato G, Merlo B, Iacono E, Bucci D, Seren E, Tamanini C, Galeati G, Spinaci M
Veterinary Clinical Department, University of Bologna, Bologna, Italy.
Reprod Domest Anim. 2010 Apr;45(2):331-5. doi: 10.1111/j.1439-0531.2008.01314.x. Epub 2008 Dec 15.
Little information is available on the quality of stallion spermatozoa after sex sorting. The objectives of the present study were to assess the quality of sex-sorted stallion spermatozoa and determine its fertilizing ability after hysteroscopic low dose insemination. Ejaculates from four stallions were collected and sorted by a MoFlo SX flow cytometer/sperm sorter. Before and after sorting, spermatozoa were evaluated for motility by Computer Assisted Sperm Analysis, viability (SYBR 14-propidium iodide), mitochondrial function (JC-1) and acrosomal status (fluorescein isothiocyanate Pisum sativum agglutinin conjugated). A fertility trial was carried out on four mares (seven oestrous cycles) by hysteroscopic insemination, depositing 5 x 10(6) X-bearing spermatozoa. Sex sorting resulted in a significant decrease (p < 0.001) in all motility characteristics. Sperm viability and percentage of spermatozoa with functional mitochondria were not affected by the sorting process, while the percentage of reacted spermatozoa was higher (p < 0.01) for non-sorted than sorted spermatozoa. Pregnancy rate was 28.6% (2/7) after low dose hysteroscopic insemination. Only one pregnancy was carried to term with the birth of a healthy filly. In conclusion, despite the reduction in sperm motility, sex sorting did not impair stallion sperm viability and mitochondrial activity immediately post-thaw; moreover, the sexed spermatozoa retained the ability to fertilize in vivo.
关于种马精子性别分选后的质量，目前可用信息较少。本研究的目的是评估性别分选后的种马精子质量，并确定其经宫腔镜低剂量授精后的受精能力。采集了四匹种马的精液，并通过MoFlo SX流式细胞仪/精子分选仪进行分选。在分选前后，通过计算机辅助精子分析评估精子活力、通过SYBR 14-碘化丙啶评估生存力、通过JC-1评估线粒体功能以及通过异硫氰酸荧光素标记的豌豆凝集素评估顶体状态。对四匹母马（七个发情周期）进行了宫腔镜授精的生育试验，植入5×10⁶个携带X染色体的精子。性别分选导致所有活力特征显著下降（p<0.001）。精子生存力和具有功能性线粒体的精子百分比不受分选过程影响，而未分选精子的反应性精子百分比高于分选精子（p<0.01）。低剂量宫腔镜授精后的妊娠率为28.6%（2/7）。只有一次妊娠足月，产下一匹健康的小母马。总之，尽管精子活力降低，但性别分选并未立即损害解冻后种马精子的生存力和线粒体活性；此外，分选后的精子保留了体内受精能力。
