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Effect of amino acids and dipeptides on accumulation of ammonia in the medium during in vitro maturation and fertilization of porcine oocytes

机译:氨基酸和二肽对猪卵母细胞体外成熟和受精过程中氨在培养基中积累的影响

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摘要

>Aim:  The present study was designed to investigate the effect of amino acids and their dipeptides on the accumulation of ammonia in the medium during in vitro maturation (IVM) and in vitro fertilization (IVF) of porcine oocytes. >Methods:  The IVM and IVF media were modified North Carolina State University‐37 solution and modified Tyrode's albumin lactate pyruvate, respectively. Porcine oocytes were matured in IVM medium containing 75–2400 µmol ammonia. Amino acids (1.0 mmol) or their dipeptides (2.0 mmol) related to the urea cycle were added individually to the IVM and IVF media containing 300 µmol ammonia. Oocyte maturation and fertilization were assessed using acetic–orcein staining, and the accumulation of ammonia in the media was measured using the indophenol method. >Results:  Percentages of metaphase II (MII) were adversely affected (P < 0.05) by ≥300 µmol concentrations of ammonia in the IVM medium. In the presence of 300 µmol ammonia in the IVM and IVF media, glutamic acid, l‐alanyl‐L‐glutamine (AlaGln), l‐glycyl‐L‐glutamine (GlyGln) and AlaGln + GlyGln showed the highest rate (P < 0.05) of MII, monospermic fertilization, and the lowest rate (P < 0.05) of ammonia accumulation in the media. >Conclusion:  AlaGln and GlyGln in IVM and IVF media were more stable and effective than the individual amino acids in reducing the accumulation of ammonia, and increased the rate of porcine oocyte MII and monospermic fertilization in vitro. (Reprod Med Biol 2007; >6: 165–170)
机译:>目的:本研究旨在研究氨基酸及其二肽对猪卵母细胞体外成熟(IVM)和体外受精(IVF)期间氨在培养基中积累的影响。 >方法:分别将IVM和IVF培养基改良为北卡罗莱纳州立大学37溶液,并改良Tyrode的白蛋白乳酸酯丙酮酸。猪卵母细胞在含有75–2400 µmol氨水的IVM培养基中成熟。将与尿素循环有关的氨基酸(1.0 mmol)或其二肽(2.0 mmol)分别添加到含有300μmol氨水的IVM和IVF培养基中。卵母细胞的成熟和受精通过醋酸-大黄素染色进行评估,培养基中氨的积累采用吲哚酚法进行测量。 >结果: IVM培养基中氨浓度≥300µmol会对中期II(MII)的百分比产生不利影响(P <0.05)。在IVM和IVF培养基中存在300μmol氨气时,谷氨酸,1-丙氨酰-L-谷氨酰胺(AlaGln),1-丙氨酰-L-谷氨酰胺(GlyGln)和AlaGln +甘氨酸Gln显示最高的发生率(P <0.05 ),单精子受精以及培养基中氨的累积率最低(P <0.05)。 >结论: IVM和IVF培养基中的AlaGln和GlyGln在减少氨气积累方面比单个氨基酸更稳定,更有效,并提高了猪卵母细胞MII和单精子受精率。 (Reprod Med Biol 2007; > 6 :165-170)

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