Bermejo-Alvarez Pablo, Lonergan Patrick, Rath Detlef, Gutiérrez-Adan Alfonso, Rizos Dimitrios
Departamento de Reproducción Animal y Conservaciónde Recursos Zoogenéticos, INIA, Carretera De la Coruña Km 5.9, Madrid 28040, Spain.
Reprod Fertil Dev. 2010;22(2):426-36. doi: 10.1071/RD09142.
Using bovine embryos generated in vitro from IVF with X-sorted, Y-sorted and unsorted spermatozoa, we compared the kinetics of male and female embryo development and gene expression between male and female blastocysts. Bovine in vitro-matured oocytes (n = 8858) were fertilised with spermatozoa from each of three different bulls (X-sorted, Y-sorted or unsorted spermatozoa depending on the experiment). The cleavage rate was assessed 24, 27, 30, 33, 36, 40, 44 and 48 h post insemination (h.p.i.) and blastocyst development was recorded on Days 6-9. The relative mRNA abundance of nine genes (GSTM3, DNTM3A, PGRMC1, TP53, BAX, COX2, IGF2R, AKR1B1 and PLAC8) was analysed in male and female Day 7 blastocysts produced with sorted and unsorted spermatozoa from one bull. Cumulative cleavage rate and blastocyst yield were significantly higher in the unsorted group compared with the X- or Y-sorted group from the same bull (P < or = 0.05). Although differences existed between bulls in terms of cleavage rate, no differences were observed in cleavage rate between X- and Y-sorted spermatozoa within a bull. The blastocyst yield was significantly higher only for Bull 3 when the Y-sorted spermatozoa were used (27.1+2.8 v. 19.1+1.4 for Y- and X-sorted spermatozoa, respectively; P < 0.05). There were no differences in the mRNA abundance of the nine genes analysed between embryos of the same sex produced with sorted or unsorted spermatozoa. However, significant differences in polyA mRNA abundance were observed between male and female blastocysts for three genes (GSTM3, DNMT3A and PGRMC1; P < or = 0.05). In conclusion, the use of sorted rather than unsorted spermatozoa in IVF significantly delays the onset of first cleavage. Differences were noted between bulls, but not between X- and Y-sorted spermatozoa, and although no differences were found in terms of the mRNA abundance of the nine genes tested between sorted and unsorted spermatozoa, sex-related differences were found in the case of three genes.
利用通过体外受精（IVF）产生的牛胚胎，这些胚胎使用经X分选、Y分选和未分选的精子，我们比较了雄性和雌性胚胎发育的动力学以及雄性和雌性囊胚之间的基因表达。将牛体外成熟的卵母细胞（n = 8858）与来自三头不同公牛的精子受精（根据实验，精子分别为X分选、Y分选或未分选）。在授精后24、27、30、33、36、40、44和48小时（h.p.i.）评估卵裂率，并在第6 - 9天记录囊胚发育情况。分析了用一头公牛的分选和未分选精子产生的第7天雄性和雌性囊胚中九个基因（GSTM3、DNTM3A、PGRMC1、TP53、BAX、COX2、IGF2R、AKR1B1和PLAC8）的相对mRNA丰度。与同一头公牛的X分选或Y分选组相比，未分选组的累积卵裂率和囊胚产量显著更高（P≤0.05）。尽管不同公牛之间在卵裂率方面存在差异，但在同一头公牛内，X分选和Y分选精子之间的卵裂率未观察到差异。仅当使用Y分选精子时，公牛3的囊胚产量显著更高（Y分选和X分选精子的囊胚产量分别为27.1 + 2.8和19.1 + 1.4；P < 0.05）。在用分选或未分选精子产生的同性胚胎之间，所分析的九个基因的mRNA丰度没有差异。然而，在三个基因（GSTM3、DNMT3A和PGRMC1；P≤0.05）的雄性和雌性囊胚之间观察到了聚腺苷酸mRNA丰度的显著差异。总之，在体外受精中使用分选而非未分选的精子会显著延迟首次卵裂的开始。不同公牛之间存在差异，但X分选和Y分选精子之间没有差异，并且尽管在分选和未分选精子之间所测试的九个基因的mRNA丰度方面未发现差异，但在三个基因的情况下发现了与性别相关的差异。
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