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Published on: May 11, 2017
Guanidinoacetic acid supplementation negatively affects bovine cumulus-oocyte complexes during in vitro maturation
Alexandria Snider1, Martim Kaps1,2, Lea Rempel1
1USDA, Agricultural Research Service, U.S. Meat Animal Research Center, Clay Center, NE 68933-0166, USA.
Context:
Methylation is an important part of oocyte maturation and early embryonic development. Altered methylation status leads to poor oocyte quality and altered embryonic development. One specific metabolite, guanidinoacetic acid (GAA), sequesters methyl-groups to make creatine, causing a methyl deficient state. Understanding how GAA influences oocyte methylation status increases knowledge of oocyte maturation in vitro.
Aims:
The objective of this study was to determine the threshold concentration of GAA supplementation negatively impacting oocyte quality. We hypothesized that supplementation of GAA reduces quality of oocytes and alters early embryonic development in a dose dependent manner.
Methods:
Immature cumulus oocyte complexes were collected from slaughterhouse ovaries and matured in vitro with GAA (0 mM, 5 mM, 10 mM or 20 mM) incorporated into the medium. Matured oocytes were randomly collected to determine microtubule distribution and transcriptomic analysis, or fertilized and developed embryos in vitro.
Key Results:
Incorporation of GAA during in vitro maturation negatively impacted oocyte quality and early embryonic development. Specifically, supplementation of 5 mM GAA reduced S-adenosyl-L-methionine production (P < 0.05), increased transcripts within the creatine pathway in oocytes (P < 0.05) and impaired chromosome arrangement (P < 0.05). Reduction in total cells was observed in blastocysts from the 5 mM and 20 mM GAA group.
Conclusions:
Incorporation of 5 mM GAA to the in vitro maturation medium negatively impacted oocyte and subsequent early embryonic development. The 20 mM supplementation demonstrated negative implications, but the methylation pathway was not impacted.
Implications:
Further research is warranted to investigate other potential mechanisms that are impacted by eliciting a methyl donor deficient status in matured oocytes.
