Amino acid supplementation affects imprinted gene transcription patterns in parthenogenetic porcine blastocysts

PLoS One. 2014 Sep 2;9(9):e106549. doi: 10.1371/journal.pone.0106549. eCollection 2014.

Abstract

To determine whether exogenous amino acids affect gene transcription patterns in parthenogenetic porcine embryos, we investigated the effects of amino acid mixtures in culture medium. Parthenogenetic embryos were cultured in PZM3 medium under four experimental conditions: 1) control (no amino acids except L-glutamine and taurine); 2) nonessential amino acids (NEAA); 3) essential amino acids (EAA); and 4) NEAA and EAA. The rate of development of embryos to the four-cell stage was not affected by treatment. However, fewer (P<0.05) embryos cultured with EAA (12.8%) reached the blastocyst stage as compared with the control group (25.6%) and NEAA group (30.3%). Based on these findings, we identified genes with altered expression in parthenogenetic embryos exposed to medium with or without EAAs. The results indicated that EAA influenced gene expression patterns, particularly those of imprinted genes (e.g., H19, IGF2R, PEG1, XIST). However, NEAAs did not affect impaired imprinted gene expressions induced by EAA. The results also showed that mechanistic target of rapamycin (MTOR) mRNA expression was significantly increased by EAA alone as compared with control cultures, and that the combined treatment with NEAA and EAA did not differ significantly from those of control cultures. Our results revealed that gene transcription levels in porcine embryos changed differentially depending on the presence of EAA or NEAA. However, the changes in the H19 mRNA observed in the parthenogenetic blastocysts expression level was not related to the DNA methylation status in the IGF2/H19 domain. The addition of exogenous amino acid mixtures affected not only early embryonic development, but also gene transcription levels, particularly those of imprinted genes. However, this study did not reveal how amino acids affect expression of imprinted genes under the culture conditions used. Further studies are thus required to fully evaluate how amino acids affect transcriptional regulation in porcine embryos.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / administration & dosage*
  • Amino Acids, Essential / administration & dosage
  • Animals
  • Blastocyst / drug effects*
  • Blastocyst / metabolism*
  • Culture Media / chemistry
  • DNA Methylation / drug effects
  • Embryonic Development / drug effects
  • Embryonic Development / genetics
  • Female
  • Gene Expression Regulation, Developmental / drug effects
  • Genomic Imprinting / drug effects*
  • Parthenogenesis / drug effects*
  • Parthenogenesis / genetics*
  • RNA, Long Noncoding / genetics
  • RNA, Long Noncoding / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Sus scrofa
  • TOR Serine-Threonine Kinases / genetics
  • Transcription, Genetic / drug effects

Substances

  • Amino Acids
  • Amino Acids, Essential
  • Culture Media
  • H19 long non-coding RNA
  • RNA, Long Noncoding
  • RNA, Messenger
  • TOR Serine-Threonine Kinases

Grants and funding

This work was supported by a grant of Next-Generation BioGreen 21 Program (No. PJ00832303) Rural Development Administration, and also by a grant of Research Program (No. 307-02) Gyeonggi-do Project, Republic of Korea. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.