title: Direct and Osmolarity-Dependent Effects of Glycine on Preimplantation Bovine Embryos.
abstract: Concentrations of glycine (Gly) in embryo culture media are often lower (~0.1 mM) than those in oviductal or uterine fluids (≥1.2 mM). The objective of this study was to determine direct and osmolarity-dependent effects of physiological concentrations of Gly on blastocyst formation and hatching, cell allocation to the trophectoderm (TE) and inner cell mass (ICM), and metabolic activity of bovine embryos. In experiment 1, zygotes were cultured with 100 or 120 mM NaCl and 0 or 1 mM Gly for the first 72 h of culture. Blastocyst formation and hatching were improved (P<0.05) when embryos were cultured with 100 compared to 120 mM NaCl. Inclusion of 1 mM Gly improved (P<0.05) blastocyst formation compared to 0 mM Gly, but this effect was only significant (P<0.05) for embryos cultured with 120 mM NaCl, suggesting bovine embryos can utilize Gly as an osmolyte. In experiment 2, embryos were cultured with 0.1, 1.1, 2.1, or 4.1 mM Gly (100 mM NaCl) for the final 96 h of culture. Blastocyst development was not affected (P>0.05) by Gly, but hatching (0.1 mM Gly, 18.2%) was improved (P<0.05) when embryos were cultured with 1.1 (31.4%) or 2.1 (29.4%) mM Gly. Blastocyst, TE, and ICM cell numbers were not affected (P>0.05) by Gly in either experiment. Blastocysts produced alanine, glutamine, pyruvate, and urea and consumed aspartate, but this metabolic profile was not affected (P>0.05) by Gly. In conclusion, Gly (1.0 mM) improves the development of both early and late stage embryos, but beneficial effects are more pronounced for early embryos exposed to elevated osmolarity.
authors.affiliation: National Foundation for Fertility Research, 10290 RidgeGate Cr., Lone Tree, CO, 80124, United States of America.
authors.forename: Jason R
authors.identifier: http://orcid.org/0000-0002-1123-0958
authors.initials: JR
authors.lastname: Herrick
authors.affiliation: Proteomics and Metabolomics Facility, Colorado State University, C-121 Microbiology Building, 2021 Campus Delivery, Fort Collins, CO, 80523, United States of America.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Lyons
authors.affiliation: National Foundation for Fertility Research, 10290 RidgeGate Cr., Lone Tree, CO, 80124, United States of America.
authors.forename: Alison F
authors.initials: AF
authors.lastname: Greene
authors.affiliation: Proteomics and Metabolomics Facility, Colorado State University, C-121 Microbiology Building, 2021 Campus Delivery, Fort Collins, CO, 80523, United States of America.
authors.forename: Corey D
authors.initials: CD
authors.lastname: Broeckling
authors.affiliation: Colorado Center for Reproductive Medicine, 10290 RidgeGate Cr., Lone Tree, CO, 80124, United States of America.
authors.forename: William B
authors.initials: WB
authors.lastname: Schoolcraft
authors.affiliation: National Foundation for Fertility Research, 10290 RidgeGate Cr., Lone Tree, CO, 80124, United States of America.
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Krisher
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D053624:Blastocyst Inner Cell Mass / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002417:Cattle; D047108:Embryonic Development / Q000187:drug effects; D005307:Fertilization in Vitro; D005998:Glycine / Q000378:metabolism / Q000494:pharmacology*; D009994:Osmolar Concentration; D014327:Trophoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015053:Zygote / Q000378:metabolism
chemical_list: D000596:Amino Acids; D005998:Glycine