title: Amino acid supplementation affects imprinted gene transcription patterns in parthenogenetic porcine blastocysts.
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.
authors.affiliation: Sooam Biotech Research Foundation, Seoul, Republic of Korea.
authors.forename: Chi-Hun
authors.initials: CH
authors.lastname: Park
authors.affiliation: Sooam Biotech Research Foundation, Seoul, Republic of Korea.
authors.forename: Young-Hee
authors.initials: YH
authors.lastname: Jeong
authors.affiliation: Sooam Biotech Research Foundation, Seoul, Republic of Korea.
authors.forename: Yeun-Ik
authors.initials: YI
authors.lastname: Jeong
authors.affiliation: Department of Animal Sciences, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, Republic of Korea.
authors.forename: Jeong-Woo
authors.initials: JW
authors.lastname: Kwon
authors.affiliation: Sooam Biotech Research Foundation, Seoul, Republic of Korea.
authors.forename: Taeyoung
authors.initials: T
authors.lastname: Shin
authors.affiliation: College of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, Republic of Korea.
authors.forename: Sang-Hwan
authors.initials: SH
authors.lastname: Hyun
authors.affiliation: College of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, Republic of Korea.
authors.forename: Eui-Bae
authors.initials: EB
authors.lastname: Jeung
authors.affiliation: Department of Animal Sciences, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk, Republic of Korea.
authors.forename: Nam-Hyung
authors.initials: NH
authors.lastname: Kim
authors.affiliation: Animal Quarantine Division, Gyeonggi-Do, Suwon, Republic of Korea.
authors.forename: Sang-Kyo
authors.initials: SK
authors.lastname: Seo
authors.affiliation: Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul, Republic of Korea.
authors.forename: Chang-Kyu
authors.initials: CK
authors.lastname: Lee
authors.affiliation: Sooam Biotech Research Foundation, Seoul, Republic of Korea.
authors.forename: Woo-Suk
authors.initials: WS
authors.lastname: Hwang
mesh_terms: D000596:Amino Acids / Q000008:administration & dosage*; D000601:Amino Acids, Essential / Q000008:administration & dosage; D000818:Animals; D001755:Blastocyst / Q000187:drug effects* / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry; D019175:DNA Methylation / Q000187:drug effects; D047108:Embryonic Development / Q000187:drug effects / Q000235:genetics; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D018392:Genomic Imprinting / Q000187:drug effects*; D010312:Parthenogenesis / Q000187:drug effects* / Q000235:genetics*; D062085:RNA, Long Noncoding / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034421:Sus scrofa; D058570:TOR Serine-Threonine Kinases / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D000596:Amino Acids; D000601:Amino Acids, Essential; D003470:Culture Media; C094335:H19 long non-coding RNA; D062085:RNA, Long Noncoding; D012333:RNA, Messenger; D058570:TOR Serine-Threonine Kinases