Effect of Dipeptides on In vitro Maturation, Fertilization and Subsequent Embryonic Development of Porcine Oocytes.

K M A Tareq,Hirotada Tsujii,Quzi Sharmin Akter,Inho Choi,M A M Yahia Khandoker

doi:10.5713/ajas.2012.12567

Abstract

The effects of amino acids and dipeptides on in vitro production of porcine embryos and accumulation of ammonia in culture medium during developmental stages were examined in this study. The maturation, fertilization and development of embryonic cultures were performed in modified Tissue culture medium (mTCM)-199 supplemented with 10% (v/v) porcine follicular fluid, modified Tyrode’s albumin lactate pyruvate (mTALP) medium, and modified North Carolina State University (mNCSU)-23 medium, respectively. In addition, amino acids and dipeptides of different concentrations and combinations were used to treat the embryos. The addition of L-alanyl-L-glutamine (AlnGln)+L-glycyl-L-glutamine (GlyGln) significantly (p<0.05) improved oocyte maturation, fertilization and the incorporation and oxidation of 14C(U)-glucose when compared to the control group and other treatment groups. Additionally, 2–4 cell, 8–16 cell, morula and blastocyst development increased significantly (p<0.05) following treatment with AlnGln+GlyGln when compared to the control group and other treatment groups, while this treatment reduced the accumulation of ammonia. Taken together, these findings suggest that treatment with AlnGln+GlyGln may play an important role in increasing the rate of porcine oocyte maturation, fertilization and embryonic development by reducing the level of accumulated ammonia measured in the culture media.

Highlights

In vitro maturation (IVM) and in vitro fertilization (IVF) of porcine oocytes have been extensively investigated
We previously reported that the accumulation of ammonia in the medium could be reduced by supplementation with seleon L-methionine (SeMet) and SeMet Vitamin-E (Tareq et al, 2012)
We previously reported that porcine oocyte maturation was adversely affected when 300 M exogenous ammonia was added to the maturation medium (Tareq et al, 2007)

Summary

Introduction

In vitro maturation (IVM) and in vitro fertilization (IVF) of porcine oocytes have been extensively investigated. Amino acids are included in the majority of culture media for both oocyte maturation and embryonic culture because they serve a variety of physiological functions including protein and nucleotide synthesis (Epstein and Smith, 1973; Alexiou and Leese, 1992), provision of energy sources (Gardner, 1998), protection against osmotic shock (Lane, 2001) with oxidative stress (Lindenbaum, 1973), and regulation of pH (Edwards et al, 1998). Spontaneous degradation and catabolism of amino acids, glutamine (Gln), can result in the production of ammonia (Gardner and Lane, 1993), which is toxic to living cells both in vivo (Prior and Visek, 1972) and in vitro (Visek et al, 1972).

Methods

Results

Conclusion