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

Abstract

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 75-2400µmol ammonia. Amino acids (1.0mmol) or their dipeptides (2.0mmol) related to the urea cycle were added individually to the IVM and IVF media containing 300µmol 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 ≥300µmol concentrations of ammonia in the IVM medium. In the presence of 300µmol 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).