Lee Ji-Eun, Hwangbo Yong, Cheong Hee-Tae, Yang Boo-Keun, Park Choon-Keun
College of Animal Life Sciences, Kangwon National University, Chunchoen 24341, Korea.
College of Veterinary Medicine, Kangwon National University, Chunchoen 24341, Korea.
Dev Reprod. 2018 Dec;22(4):297-307. doi: 10.12717/DR.2018.22.4.297. Epub 2018 Dec 31.
The objective of this study was to evaluate the effects of alpha-linolenic acid (ALA) during maturation (IVM) on cumulus expansion, nuclear maturation, fertilization capacity and subsequent development in porcine oocytes. The oocytes were incubated with 0, 25, 50, and 100 μM ALA. Cumulus expansion was measured at 22 h, and gene expresison and nuclear maturation were analyzed at 44 h after maturation. Then, mature oocytes with ALA were inseminated, and fertilization parameters and embryo development were evaluated. In results, both of cumulus expansion and nuclear maturation were increased in 50 μM ALA groups compared to control groups (<0.05). However, expression of gap junction protein alpha 1 (GJA1, cumulus expansion-related gene), delta-6 desaturase (FADS1, fatty acid metabolism-related gene), and delta-5 desaturase () mRNA in cumulus cells were reduced by 50 μM ALA treatment (<0.05). Cleavage rate was enhanced in 25 and 50 μM ALA groups (<0.05), especially, treatment of 50 μM ALA promoted early embryo develop to 4 and 8 cell stages (<0.05). However, blastocyst formation and number of cells in blastocyst were not differ in 25 and 50 μM ALA groups. Our findings show that ALA treatment during maturation could improve nuclear maturation, fertilization, and early embryo development through enhancing of cumulus expansion, however, fatty acid metabolism- and cumulus expansion-related genes were down-regulated. Therefore, addition of ALA during IVM of oocytes could improve fertilization and developmental competence, and further studies regarding with the mechanism of ALA metabolism are needed.
本研究的目的是评估α-亚麻酸(ALA)在猪卵母细胞体外成熟(IVM)过程中对卵丘扩展、核成熟、受精能力及后续发育的影响。将卵母细胞分别与0、25、50和100μM的ALA共同孵育。在22小时时测量卵丘扩展情况,并在成熟后44小时分析基因表达和核成熟情况。然后,对添加ALA的成熟卵母细胞进行授精,并评估受精参数和胚胎发育情况。结果显示,与对照组相比,50μM ALA组的卵丘扩展和核成熟均有所增加(<0.05)。然而,50μM ALA处理使卵丘细胞中缝隙连接蛋白α1(GJA1,与卵丘扩展相关基因)\u3001Δ-6去饱和酶(FADS1,脂肪酸代谢相关基因)和Δ-5去饱和酶()mRNA的表达降低(<0.05)。25和50μM ALA组的卵裂率有所提高(<0.05),特别是50μM ALA处理促进早期胚胎发育至4细胞和8细胞阶段(<0.05)。然而,25和50μM ALA组的囊胚形成率和囊胚细胞数并无差异。我们的研究结果表明,成熟过程中ALA处理可通过增强卵丘扩展来改善核成熟、受精和早期胚胎发育,然而,脂肪酸代谢和卵丘扩展相关基因表达下调。因此,卵母细胞IVM过程中添加ALA可提高受精和发育能力,关于ALA代谢机制还需要进一步研究。