Role of nonessential amino acids on porcine embryos produced by parthenogenesis or somatic cell nuclear transfer.
Amino acids play a multitude of roles during early embryonic development and have been demonstrated to facilitate improved development of in vivo or in vitro fertilized and parthenogenetic embryos in several species. However, review of emerging literatures, shows that culture milieu of cloned embryos might be different from those of in vitro fertilized embryos. This study therefore, evaluated the effect of nonessential amino acids (NEAA) on yield and quality of porcine embryos produced by somatic cell nuclear transfer (SCNT) and compared them with parthenogenetic embryos as control. Analysis showed that, supplementation of NEAA to culture medium significantly improved the blastocyst rate of parthenogenetic (38.9 +/- 8.8 vs. 27.5 +/- 9.0%) as well as SCNT (22.5 +/- 2.2 vs. 13.8 +/- 3.4%) embryos although cleavage rates were not different (P < 0.05). These blastocysts also had higher hatching ability and contained higher cell number than control blastocysts (P < 0.05). TUNEL labeling revealed that blastocysts cultured in the presence of NEAA were less predisposed to biochemical apoptosis and showed lower indices of TUNEL, fragmentation, and total apoptosis than those cultured in the absence of NEAA (P < 0.05). Real-time qRT-PCR analysis further revealed that NEAA decreased the expression ratio of BAX:BCL-xL and enhanced the relative abundance of IGF2 transcripts. Therefore, our study suggests that NEAA improves the yield and quality of cloned porcine embryos by enhancing blastocyst expansion, hatching, and total cell number and decreasing the apoptosis by positively modulating the expression of embryo survival related genes, similar to those reported for in vivo or in vitro fertilized embryos. Nonessential amino acids improve the yield and quality of cloned and parthenogenetic porcine embryos and modulate the expression of embryo survival related genes.
Gupta
Mukesh Kumar
MK
Department of Animal Biotechnology, Bio-Organ Research Center, Konkuk University, 1, Hwayang-dong, Gwangjin-Gu, Seoul, South Korea.
Uhm
Sang Jun
SJ
Lee
Sung-Ho
SH
Lee
Hoon Taek
HT
D000596:Amino Acids / Q000494:pharmacology*; D000818:Animals; D046149:Embryo Culture Techniques / Q000379:methods; D047108:Embryonic Development / Q000187:drug effects*; D053595:Embryonic Stem Cells / Q000187:drug effects* / Q000378:metabolism; D020869:Gene Expression Profiling; D007335:Insulin-Like Growth Factor II / Q000187:drug effects / Q000235:genetics; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010312:Parthenogenesis / Q000187:drug effects*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D051028:bcl-2-Associated X Protein / Q000187:drug effects / Q000235:genetics; D051020:bcl-X Protein / Q000187:drug effects / Q000235:genetics
D000596:Amino Acids; D051028:bcl-2-Associated X Protein; D051020:bcl-X Protein; D007335:Insulin-Like Growth Factor II