Madrid-Bury Ninoska, Fernández Raúl, Jiménez Adela, Pérez-Garnelo Sonia, Moreira Pedro Nuno, Pintado Belén, de la Fuente Julio, Gutiérrez-Adán Alfonso
Departamento de Reproducción Animal y Conservación de Recursos Zoogenéticos, INIA, Ctra de la Coruña Km 5.9, Madrid 28040, Spain.
Zygote. 2003 Aug;11(3):229-35. doi: 10.1017/s0967199403002272.
Offspring gender preselection has applications of considerable economic, health and ecological interest. In this study we analysed modifications of the percentages of spermatozoa bearing Y and X chromosomes when semen samples are submitted to a double swim-up technique as a possible method for producing embryos of known sex with in vitro fertilisation protocols. As an initial experiment to provide accurate evaluation of the method we determined the possible incidence of natural deviations in the primary sex ratio between bulls or ejaculates, analysing the percentage of Y-chromosome DNA bearing spermatozoa (%Y-CDBS) with a polymerase chain reaction (PCR) amplification of X- and Y-specific fragments. Ejaculates were tested by direct semiquantitative PCR sexing and by sexing blastocysts produced in vitro with these spermatozoa. Bulls and ejaculates did not have any effect on the %Y-CDBS or on the sex ratio of embryos produced in vitro using these ejaculates. However, our double swim-up sperm preparation method produced differences in %Y-CDBS in some of the sperm fractions, suggesting that there are intrinsic differences in capacitation of X- and Y-bearing spermatozoa that might be used to produce embryos of the desired sex with in vitro fertilisation.
后代性别预选在经济、健康和生态方面具有重要应用价值。在本研究中，我们分析了精液样本采用双重上浮技术处理时，携带Y和X染色体精子百分比的变化情况，以此作为一种在体外受精方案中生产已知性别的胚胎的可能方法。作为对该方法进行准确评估的初步实验，我们通过聚合酶链反应（PCR）扩增X和Y特异性片段来分析Y染色体DNA携带精子的百分比（%Y-CDBS），从而确定公牛或射精之间原发性别比例自然偏差的可能发生率。通过直接半定量PCR性别鉴定以及对使用这些精子体外产生的囊胚进行性别鉴定来检测射精样本。公牛和射精样本对%Y-CDBS或使用这些射精样本体外产生的胚胎性别比例均无影响。然而，我们的双重上浮精子制备方法在某些精子组分中产生了%Y-CDBS的差异，这表明携带X和Y的精子在获能方面存在内在差异，这种差异可能用于在体外受精时生产所需性别的胚胎。
