Supplementation of culture medium with L-carnitine improves development and cryotolerance of bovine embryos produced in vitro
Toshikiyo Takahashi A B D , Yasushi Inaba A D , Tamas Somfai A E , Masahiro Kaneda A , Masaya Geshi A , Takashi Nagai A and Noboru Manabe C
+ Author Affiliations
- Author Affiliations
A NARO Institute of Livestock and Grassland Science, Ikenodai 2, Tsukuba, Ibaraki 305-0901, Japan.
B Livestock Experiment Station, Akita Prefectural Agriculture Forestry and Fisheries Research Centre, 13-3 Kaiso-numayachi, Jinguji, Daisen, Akita 019-1701, Japan.
C Animal Resource Science Centre, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Ago 3145, Kasama, Ibaraki 319-0206, Japan.
D These authors contributed equally to the paper.
E Corresponding author. Email: somfai@affrc.go.jp
Reproduction, Fertility and Development 25(4) 589-599 https://doi.org/10.1071/RD11262
Submitted: 12 October 2011 Accepted: 4 May 2012 Published: 4 June 2012
Abstract
High lipid content in embryos is associated with low freezing tolerance. This study assessed the effects of exogenous l-carnitine, an enhancer of lipid metabolism, on the in vitro development and freezing survival of bovine embryos. Also, effects on metabolic activity, reactive oxygen species (ROS) and apoptosis were investigated. Supplementation of embryo culture medium with 1.518 mM or 3.030 mM l-carnitine significantly increased the rates of zygote development to the blastocyst stage and blastocyst cell numbers whereas 6.072 mM of this compound did not improve embryo development. Survival rates after slow freezing of blastocysts were significantly higher when embryos were cultured in the presence of 1.518 mM or 3.030 mM l-carnitine compared with the control. A lower density of lipid droplets was detected in l-carnitine-treated blastocysts compared with the control. l-carnitine significantly reduced ROS levels in 2-cell embryos but did not reduce ROS levels at later stages. The apoptotic cell rate was not different between control and l-carnitine-treated blastocysts. l-carnitine significantly increased ATP levels in 2-cell embryos but not at the 8-cell or blastocyst stages. l-carnitine increased the expression of metabolism-related ATP6 and COX1 genes in blastocysts. In conclusion, l-carnitine supplementation enhanced lipid metabolism in embryos resulting in improved development and cryotolerance of bovine blastocysts produced in vitro.
Additional keywords: freezing, lipid, metabolism.
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