Bermejo-Alvarez P, Rizos D, Rath D, Lonergan P, Gutiérrez-Adán A
Departamento de Reproducción Animal y Conservación de Recursos Zoogenéticos, INIA, 28040 Madrid, Spain.
Biol Reprod. 2008 Oct;79(4):594-7. doi: 10.1095/biolreprod.108.070169. Epub 2008 Jun 25.
It has been reported that the mammalian female could have a preconceptual influence on the sex of her offspring, and it has been hypothesized that this influence could go some way toward accounting for the reported lower fertility following insemination with sex-sorted sperm. To test whether in vitro matured oocytes are able to select X- or Y-bearing spermatozoa following in vitro fertilization (IVF), we fertilized in vitro 1788 oocytes with X-sorted semen, Y-sorted semen, a mix of X- and Y-sorted semen, and unsorted semen from the same bull, and cultured until Day 9. Fertility was assessed by recording cleavage rate at 48 h postinsemination (hpi) and blastocyst development until Day 9. Embryos were sexed at the two- to four-cell stage and the blastocyst stage. The proportion of zygotes cleaving at 48 hpi was not different between X- and Y-sorted groups and the mix of X- and Y-sorted semen group; however, all were significantly lower than the unsorted group (P < 0.001). Blastocyst yield on Day 6 was significantly higher (P < or = 0.01) in the control group compared with the rest of the groups. Cumulative blastocyst yields on Days 7, 8, and 9 were also significantly higher (P < or = 0.01) in the unsorted group compared with the sorted groups. The proportion of female and male two- to four-cell embryos obtained following IVF with X- and Y-sorted sperm was 88% and 89%, respectively and the sex ratio at the two- to four-cell stage was not different following IVF with unsorted or sorted/recombined sperm (56.9% males vs. 57% males, respectively). At the blastocyst stage, similar percentages were obtained. In conclusion, the differences in cleavage and blastocyst development using sorted versus unsorted sperm are not due to the oocyte preferentially selecting sperm of one sex over another, but are more likely due to spermatic damage caused by the sorting procedure.
据报道,哺乳动物雌性可能对其后代的性别有受孕前的影响,并且有人推测这种影响在一定程度上可以解释在用性别分选精子授精后所报道的较低生育力。为了测试体外成熟的卵母细胞在体外受精(IVF)后是否能够选择携带X或Y的精子,我们用X分选精液、Y分选精液、X和Y分选精液的混合物以及来自同一头公牛的未分选精液对1788个卵母细胞进行体外受精,并培养至第9天。通过记录授精后48小时(hpi)的卵裂率和直到第9天的囊胚发育来评估生育力。在二至四细胞阶段和囊胚阶段对胚胎进行性别鉴定。X分选组、Y分选组以及X和Y分选精液混合组在48 hpi时的受精卵分裂比例没有差异;然而,所有这些组均显著低于未分选组(P < 0.001)。与其他组相比,对照组在第6天的囊胚产量显著更高(P ≤ 0.01)。与分选组相比,未分选组在第7、8和9天的累积囊胚产量也显著更高(P ≤ 0.01)。用X和Y分选精子进行IVF后获得的雌性和雄性二至四细胞胚胎的比例分别为88%和89%,并且用未分选或分选/重组精子进行IVF后,二至四细胞阶段的性别比例没有差异(分别为56.9%的雄性对57%的雄性)。在囊胚阶段,获得了相似的百分比。总之,使用分选精子与未分选精子在卵裂和囊胚发育上的差异不是由于卵母细胞优先选择一种性别的精子,而更可能是由于分选过程导致的精子损伤。