Department of Reproductive Sciences, Center for Species Survival, Smithsonian Conservation Biology Institute, Front Royal, Virginia 22630, USA.
Mol Reprod Dev. 2012 Mar;79(3):186-96. doi: 10.1002/mrd.22011. Epub 2011 Dec 27.
We tested the hypothesis that meiotic competence of dog oocytes is tightly linked with donor follicle size and energy metabolism. Oocytes were recovered from small (<1 mm diameter, n = 327), medium (1-<2 mm, n = 292) or large (≥2 mm, n = 102) follicles, cultured for 0, 24, or 48 hr, and then assessed for glycolysis, glucose oxidation, pyruvate uptake, glutamine oxidation, and nuclear status. More oocytes (P < 0.05) from large follicles (37%) reached the metaphase-II (MII) stage than from the small group (11%), with the medium-sized class being intermediate (18%; P > 0.05). Glycolytic rate increased (P < 0.05) as oocytes progressed from the germinal vesicle (GV) to MII stage. After 48 hr of culture, oocytes completing nuclear maturation had higher (P < 0.05) glycolytic rates than those arrested at earlier stages. GV oocytes recovered from large follicle oocytes had higher (P < 0.05) metabolism than those from smaller counterparts at culture onset. MII oocytes from large follicles oxidized more (P < 0.05) glutamine than the same stage gametes recovered from smaller counterparts. In summary, larger-sized dog follicles contain a more metabolically active oocyte with a greater chance of achieving nuclear maturation in vitro. These findings demonstrate a significant role for energy metabolism in promoting dog oocyte maturation, information that will be useful for improving culture systems for rescuing intraovarian genetic material.
我们检验了这样一个假设,即卵母细胞的减数分裂能力与供体卵泡的大小和能量代谢密切相关。从小(<1mm 直径,n=327)、中(1-<2mm,n=292)或大(≥2mm,n=102)卵泡中回收卵母细胞,分别培养 0、24 或 48 小时,然后评估糖酵解、葡萄糖氧化、丙酮酸摄取、谷氨酰胺氧化和核状态。与小卵泡(11%)相比,大卵泡(37%)中的更多卵母细胞(P<0.05)达到中期 II(MII)阶段,中卵泡组处于中间水平(18%;P>0.05)。随着卵母细胞从生发泡期(GV)向 MII 期的发展,糖酵解率增加(P<0.05)。培养 48 小时后,完成核成熟的卵母细胞的糖酵解率高于早期停滞的卵母细胞(P<0.05)。大卵泡卵母细胞回收的 GV 期卵母细胞在培养开始时的代谢率高于小卵泡卵母细胞(P<0.05)。大卵泡的 MII 期卵母细胞比从小卵泡回收的相同阶段的配子氧化更多(P<0.05)的谷氨酰胺。总之,较大的狗卵泡含有更具代谢活性的卵母细胞,在体外实现核成熟的机会更大。这些发现表明能量代谢在促进狗卵母细胞成熟中起着重要作用,这些信息将有助于改善用于挽救卵巢内遗传物质的培养系统。