Lowe Jenna L, Bartolac Louise K, Bathgate Roslyn, Grupen Christopher G
Faculty of Veterinary Science, School of Life and Environmental Sciences, The University of Sydney, NSW 2570, Australia.
J Reprod Dev. 2017 Jun 21;63(3):263-270. doi: 10.1262/jrd.2016-141. Epub 2017 Mar 16.
Sufficient generation of adenosine triphosphate (ATP) by oocytes is critical for fertilization and embryo development. The objective of this study was to determine the effects of supplementing media with L-carnitine, a co-factor required for the metabolism of fatty acids, during the peri-fertilization period on embryo development and energy generation. Firstly, in vitro matured (IVM) porcine oocytes were co-incubated with sperm in IVF medium supplemented with 0‒24 mM L-carnitine. The blastocyst formation rate of the control group was greater than those of the L-carnitine groups (P < 0.05), except for the 3 mM L-carnitine group. Subsequently, oocytes and/or sperm were treated without or with 3 mM L-carnitine for either the 1 h pre-IVF oocyte incubation; the pre-IVF sperm preparation; the first 30 min of IVF; or the entire 5.5 h of IVF. Despite similar fertilization rates among the groups, the cleavage rate of the pre-IVF oocyte group was significantly greater than those of the other groups, except for the pre-IVF sperm group. Additionally, the oocyte ATP content and the cryotolerance of the resulting blastocysts were examined following the pre-IVF oocyte treatment. Oocyte ATP content was also similar among the groups (P > 0.05). Following vitrification, the post-warming survival rate of blastocysts derived from L-carnitine-treated oocytes was greater than that of blastocysts derived from untreated oocytes (42.4% vs. 24.9%; P < 0.05). In conclusion, a 1 h oocyte exposure to 3 mM L-carnitine immediately prior to insemination enhanced cleavage and improved the cryotolerance of resulting blastocysts. While the findings are suggestive of a lipolytic action, further studies are required to clarify the contributions of lipid metabolism and oxidative mechanisms to the observed effects of the L-carnitine treatment.
卵母细胞充分生成三磷酸腺苷(ATP)对于受精和胚胎发育至关重要。本研究的目的是确定在受精前后时期向培养基中添加L-肉碱(脂肪酸代谢所需的一种辅助因子)对胚胎发育和能量生成的影响。首先,将体外成熟(IVM)的猪卵母细胞与精子在添加了0至24 mM L-肉碱的体外受精(IVF)培养基中共同孵育。对照组的囊胚形成率高于L-肉碱组(P < 0.05),3 mM L-肉碱组除外。随后,对卵母细胞和/或精子在IVF前1小时的卵母细胞孵育、IVF前的精子制备、IVF的前30分钟或整个5.5小时的IVF过程中进行不添加或添加3 mM L-肉碱的处理。尽管各组间受精率相似,但IVF前卵母细胞组的卵裂率显著高于其他组,IVF前精子组除外。此外,在IVF前卵母细胞处理后,检测了卵母细胞的ATP含量以及所得囊胚的耐冻性。各组间卵母细胞ATP含量也相似(P > 0.05)。玻璃化冷冻后,来自L-肉碱处理卵母细胞的囊胚解冻后存活率高于未处理卵母细胞的囊胚(42.4%对24.9%;P < 0.05)。总之,授精前1小时让卵母细胞暴露于3 mM L-肉碱可提高卵裂率并改善所得囊胚的耐冻性。虽然这些发现提示了脂解作用,但需要进一步研究来阐明脂质代谢和氧化机制对L-肉碱处理所观察到的效果的贡献。