Reproductive and Developmental Biology Laboratory, Department of Health, Animal Science and Food Safety, University of Milan, Milan, Italy.
Biol Reprod. 2013 Mar 7;88(3):58. doi: 10.1095/biolreprod.112.106856. Print 2013 Mar.
Oocytes isolated from cows of reproductive age with reduced antral follicle counts (AFC) have a diminished capacity of embryonic development, which may be related to alterations in the mechanism that directs the proper segregation of chromosomes. Because we demonstrated that progesterone receptor membrane component 1 (PGRMC1) is involved in chromosome congression and metaphase II (MII) plate formation, the present study was designed to determine 1) if the decrease in oocyte developmental competence observed in dairy cows with a reduced AFC is due to a higher incidence of aneuploidy and 2) whether alterations in PGRMC1 contributes to the incidence of aneuploidy. Oocytes from ovaries with reduced AFC and age-matched controls were matured in vitro and the occurrence of aneuploidy determined as well as the mRNA level and localization of PGRMC1. Although oocytes from ovaries with reduced AFC were capable of undergoing meiosis in vitro, these oocytes showed a 3-fold increase in aneuploidy compared to oocytes isolated from control ovaries (P < 0.05). Although Pgrmc1 mRNA levels were not altered, PGRMC1 and aurora kinase B (AURKB) failed to localize to precise focal points on MII chromosomes of oocytes from ovaries with reduced AFC. Furthermore, when oocytes of control ovaries were cultured with an inhibitor of AURKB activity, their MII plate was disrupted and PGRMC1 was not properly localized to the chromosomes. These results suggest that alterations in PGRMC1 and/or AURKB localization account in part for the increased aneuploidy and low development competence of oocytes from ovaries with reduced AFC.
从卵巢储备功能降低(AFC)的适龄奶牛中分离的卵母细胞胚胎发育能力下降,这可能与指导染色体正确分离的机制改变有关。因为我们已经证明孕激素受体膜成分 1(PGRMC1)参与染色体聚合并形成中期 II(MII)板,所以本研究旨在确定 1)在 AFC 减少的奶牛中观察到的卵母细胞发育能力下降是否是由于非整倍体发生率增加引起的,以及 2)PGRMC1 的改变是否导致非整倍体的发生。从 AFC 减少和年龄匹配的对照组卵巢中分离的卵母细胞在体外成熟,并确定非整倍体的发生情况,以及 PGRMC1 的 mRNA 水平和定位。尽管 AFC 减少的卵巢中的卵母细胞能够在体外进行减数分裂,但与从对照组卵巢中分离的卵母细胞相比,这些卵母细胞的非整倍体发生率增加了 3 倍(P < 0.05)。尽管 Pgrmc1 mRNA 水平没有改变,但 AFC 减少的卵巢中的卵母细胞的 PGRMC1 和极光激酶 B(AURKB)未能定位于 MII 染色体的精确焦点上。此外,当对照组卵巢中的卵母细胞用 AURKB 活性抑制剂培养时,其 MII 板被破坏,PGRMC1 不能正确定位于染色体上。这些结果表明,PGRMC1 和/或 AURKB 定位的改变部分解释了 AFC 减少的卵巢中卵母细胞非整倍体增加和发育能力下降的原因。