title: Glycine supplementation in vitro enhances porcine preimplantation embryo cell number and decreases apoptosis but does not lead to live births.
abstract: Most in vitro culture conditions are less-than-optimal for embryo development. Here, we used a transcriptional-profiling database to identify culture-induced differences in gene expression in porcine blastocysts compared to in vivo-produced counterparts. Genes involved in glycine transport (SLC6A9), glycine metabolism (GLDC, GCSH, DLD, and AMT), and serine metabolism (PSAT1, PSPH, and PHGDH) were differentially expressed. Addition of 10 mM glycine to the culture medium (currently containing 0.1 mM) reduced the abundance of SLC6A9 transcript and increased total cell number, primarily in the trophectoderm lineage (P = 0.003); this was likely by decreasing the percentage of apoptotic nuclei. As serine and glycine can be reversibly metabolized by serine hydroxymethyltransferase 2 (SHMT2), we assessed the abundance of SHMT2 transcript as well as its functional role by inhibiting it with aminomethylphosphonic acid (AMPA), a glycine analog, during in vitro culture. Both AMPA supplementation and elevated glycine decreased the mRNA abundance of SHMT2 and tumor protein p53 (TP53), which is activated in response to cellular stress, compared to controls (P ≤ 0.02). On the other hand, mitochondrial activity of blastocysts, mtDNA copy number, and abundance of mitochondria-related transcripts did not differ between control and 10 mM glycine culture conditions. Despite improvements to these metrics of blastocyst quality, transfer of embryos cultured in 10 mM glycine did not result in pregnancy whereas the transfer of in vitro-produced embryos cultured in control medium yielded live births. Mol. Reprod. Dev. 83: 246-258, 2016. © 2016 The Authors.
authors.affiliation: Division of Animal Sciences, Animal Science Research Center, Columbia, Missouri.
authors.forename: Bethany K
authors.initials: BK
authors.lastname: Redel
authors.affiliation: Division of Animal Sciences, Animal Science Research Center, Columbia, Missouri.
authors.forename: Lee D
authors.initials: LD
authors.lastname: Spate
authors.affiliation: Division of Animal Sciences, Animal Science Research Center, Columbia, Missouri.
authors.forename: Kiho
authors.initials: K
authors.lastname: Lee
authors.affiliation: Division of Animal Sciences, Animal Science Research Center, Columbia, Missouri.
authors.forename: Jiude
authors.initials: J
authors.lastname: Mao
authors.affiliation: Division of Animal Sciences, Animal Science Research Center, Columbia, Missouri.
authors.forename: Kristin M
authors.initials: KM
authors.lastname: Whitworth
authors.affiliation: Division of Animal Sciences, Animal Science Research Center, Columbia, Missouri.
authors.forename: Randall S
authors.initials: RS
authors.lastname: Prather
mesh_terms: D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D001755:Blastocyst / Q000378:metabolism*; D004624:Embryo Transfer; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D005998:Glycine / Q000494:pharmacology*; D011247:Pregnancy; D013552:Swine
chemical_list: D005998:Glycine
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