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氨基酸和二肽对猪卵母细胞成熟和受精过程中培养基中氨积累的影响。

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

作者信息

Tareq Kma, Miah Abdul Gaffar, Salma Ummay, Yoshida Mayumi, Tsujii Hirotada

机构信息

Laboratory of Animal Biotechnology, Interdisciplinary Graduate School of Science and Technology, Shinshu University, Nagano, Japan.

出版信息

Reprod Med Biol. 2007 Aug 6;6(3):165-170. doi: 10.1111/j.1447-0578.2007.00180.x. eCollection 2007 Sep.

Abstract

The present study was designed to investigate the effect of amino acids and their dipeptides on the accumulation of ammonia in the medium during maturation (IVM) and fertilization (IVF) of porcine oocytes. 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. Percentages of metaphase II (MII) were adversely affected ( 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 ( 0.05) of MII, monospermic fertilization, and the lowest rate ( 0.05) of ammonia accumulation in the media. 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 . (Reprod Med Biol 2007; : 165-170).

摘要

本研究旨在探讨氨基酸及其二肽对猪卵母细胞体外成熟(IVM)和体外受精(IVF)过程中培养基中氨积累的影响。IVM和IVF培养基分别为改良的北卡罗来纳州立大学-37溶液和改良的泰勒氏白蛋白乳酸丙酮酸溶液。猪卵母细胞在含有75 - 2400µmol氨的IVM培养基中成熟。将与尿素循环相关的氨基酸(1.0 mmol)或其二肽(2.0 mmol)分别添加到含有300µmol氨的IVM和IVF培养基中。使用醋酸洋红染色评估卵母细胞的成熟和受精情况,并使用靛酚法测量培养基中氨的积累。IVM培养基中氨浓度≥300µmol时,中期II(MII)的百分比受到不利影响(P<0.05)。在IVM和IVF培养基中存在300µmol氨的情况下,谷氨酸、L-丙氨酰-L-谷氨酰胺(AlaGln)、L-甘氨酰-L-谷氨酰胺(GlyGln)以及AlaGln + GlyGln显示出最高的MII率、单精子受精率(P<0.05),且培养基中氨积累率最低(P<0.05)。IVM和IVF培养基中的AlaGln和GlyGln在减少氨积累方面比单个氨基酸更稳定、更有效,并且提高了猪卵母细胞MII率和单精子受精率。(《生殖医学与生物学》2007年;[具体卷期]:165 - 170)

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