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31 May 2018 Glutamine supplementation enhances development of in vitro-produced porcine embryos and increases leucine consumption from the medium
Paula R. Chen, Bethany K. Redel, Lee D. Spate, Tieming Ji, Shirley Rojas Salazar, Randall S. Prather
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Abstract

Improper composition of culture medium contributes to reduced viability of in vitro-produced embryos. Glutamine (Gln) is a crucial amino acid for preimplantation embryos as it supports proliferation and is involved in many different biosynthetic pathways. Previous transcriptional profiling revealed several upregulated genes related to Gln transport and metabolism in in vitro-produced porcine blastocysts compared to in vivo-produced counterparts, indicating a potential deficiency in the culture medium. Therefore, the objective of this study was to determine the effects of Gln supplementation on in vitro-produced porcine embryo development, gene expression, and metabolism. Cleaved embryos were selected and cultured in MU2 medium supplemented with 1 mM Gln (control), 3.75 mM Gln (+Gln), 3.75 mM GlutaMAX (+Max), or 3.75 mM alanine (+Ala) until day 6. Embryos cultured with +Gln or +Max had increased development to the blastocyst stage and total number of nuclei compared to the control (P < 0.05). Moreover, expression of misregulated transcripts involved in glutamine and glutamate transport and metabolism was corrected when embryos were cultured with+Gln or+Max. Metabolomics analysis revealed increased production of glutamine and glutamate into the medium by embryos cultured with +Max and increased consumption of leucine by embryos cultured with +Gln or +Max. As an indicator of cellular health, mitochondrialmembrane potential was increased when embryos were cultured with +Max which was coincident with decreased apoptosis in these blastocysts. Lastly, two embryo transfers by using embryos cultured with +Max resulted in viable piglets, confirming that this treatment is consistent with in vivo developmental competence.

Summary Sentence

Supplemental glutamine in porcine embryo culture medium modulates gene expression and enhances metabolic activity of in vitro-produced blastocysts.

© The Author(s) 2018. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com
Paula R. Chen, Bethany K. Redel, Lee D. Spate, Tieming Ji, Shirley Rojas Salazar, and Randall S. Prather "Glutamine supplementation enhances development of in vitro-produced porcine embryos and increases leucine consumption from the medium," Biology of Reproduction 99(5), 938-948, (31 May 2018). https://doi.org/10.1093/biolre/ioy129
Received: 13 April 2018; Accepted: 30 May 2018; Published: 31 May 2018
JOURNAL ARTICLE
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KEYWORDS
Apoptosis
blastocyst
embryo culture
gene expression
metabolism
mitochondria
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