Register      Login
Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Enhancement of lipid metabolism with L-carnitine during in vitro maturation improves nuclear maturation and cleavage ability of follicular porcine oocytes

Tamás Somfai A D E , Masahiro Kaneda A , Satoshi Akagi A , Shinya Watanabe A , Seiki Haraguchi A , Eiji Mizutani A , Thanh Quang Dang-Nguyen B , Masaya Geshi A , Kazuhiro Kikuchi C and Takashi Nagai A
+ Author Affiliations
- Author Affiliations

A National Institute of Livestock and Grassland Science, National Agriculture and Food Research Organisation, Tsukuba, Ibaraki 305-0901, Japan.

B Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8577, Japan.

C National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan.

D Animal Breeding and Reproduction Division, National Institute of Livestock and Grassland Science, National Agriculture and Food Research Organisation, 2 Ikenodai, Tsukuba, Ibaraki 905-0901, Japan.

E Corresponding author. Email: somfai@affrc.go.jp

Reproduction, Fertility and Development 23(7) 912-920 https://doi.org/10.1071/RD10339
Submitted: 17 December 2010  Accepted: 11 April 2011   Published: 26 August 2011

Abstract

The aim of the present study was to assess the effects of L-carnitine, an enhancer of lipid metabolism and mitochondrial activity, during in vitro maturation (IVM) on nuclear maturation and in vitro fertilisation of porcine follicular oocytes and subsequent embryo development. Mitochondrial functions, intracellular lipid content and reactive oxygen species (ROS) levels in oocytes were also investigated. L-carnitine supplementation in 0.6–5 mg mL–1 concentration during IVM significantly improved (P < 0.05) the rates of metaphase-II (MII) stage oocytes compared with the control; however, fertilisation rates and monospermy were not improved. Although supplementation of IVM medium with L-carnitine significantly increased oocyte cleavage (P < 0.05), further development to the blastocyst stage was not improved. The density of active mitochondria was significantly higher and the density of lipid droplets was significantly lower (P < 0.05) in L-carnitine-treated oocytes compared with the control. Furthermore, the ROS levels in L-carnitine-treated oocytes were significantly lower than those in the control. In conclusion, enhancing mitochondrial functions by L-carnitine improved oocyte maturation and cleavage underlining the importance of lipid metabolism for nuclear and cytoplasmic maturation of porcine oocytes.

Additional keyword: mitochondria.


References

Abdelrazik, H., Sharma, R., Mahfouz, R., and Agarwal, A. (2009). L-carnitine decreases DNA damage and improves the in vitro blastocyst development rate in mouse embryos. Fertil. Steril. 91, 589–596.
L-carnitine decreases DNA damage and improves the in vitro blastocyst development rate in mouse embryos.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXnsF2nsbc%3D&md5=341f50cd826e29c8e705563bcd85ac1aCAS | 18249387PubMed |

Abramoff, M. D., Magelhaes, P. J., and Ram, S. J. (2004). Image processing with ImageJ. Biophoton. Int. 11, 36–42.

Brevini, T. A., Vassena, R., Francisci, C., and Gandolfi, F. (2005). Role of adenosine triphosphate, active mitochondria, and microtubules in the acquisition of developmental competence of parthenogenetically activated pig oocytes. Biol. Reprod. 72, 1218–1223.
Role of adenosine triphosphate, active mitochondria, and microtubules in the acquisition of developmental competence of parthenogenetically activated pig oocytes.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXjslSntrc%3D&md5=f1cecc794620246e1a8f6c249a2d0019CAS | 15659704PubMed |

Choe, C., Shin, Y. W., Kim, E. J., Cho, S. R., Kim, H. J., Choi, S. H., Han, M. H., Han, J., Son, D. S., and Kang, D. (2010). Synergistic effects of glutathione and β-mercaptoethanol treatment during in vitro maturation of porcine oocytes on early embryonic development in a culture system supplemented with L-cysteine. J. Reprod. Dev. 56, 575–582.
Synergistic effects of glutathione and β-mercaptoethanol treatment during in vitro maturation of porcine oocytes on early embryonic development in a culture system supplemented with L-cysteine.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXhslOhsro%3D&md5=88b42feff6ba4e6908e428508a72d482CAS | 20657156PubMed |

Dunning, K. R., Cashman, K., Russell, D. L., Thompson, J. G., Norman, R. J., and Robker, R. L. (2010). Beta-oxidation is essential for mouse oocyte developmental competence and early embryo development. Biol. Reprod. 83, 909–918.
Beta-oxidation is essential for mouse oocyte developmental competence and early embryo development.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXhsFahurfN&md5=1ccb40aa6e9f3c498d9df62ea3805fefCAS | 20686180PubMed |

Genicot, G., Leroy, J. L. M. R., Van Soom, A., and Donnay, I. (2005). The use of a fluorescent dye, Nile red, to evaluate the lipid content of single mammalian oocytes. Theriogenology 63, 1181–1194.
The use of a fluorescent dye, Nile red, to evaluate the lipid content of single mammalian oocytes.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXhtlSltb8%3D&md5=01149f1052c7bd68f084ea2e45d21d68CAS | 15710202PubMed |

Hashimoto, S. (2009). Application of in vitro maturation to assisted reproductive technology. J. Reprod. Dev. 55, 1–10.
Application of in vitro maturation to assisted reproductive technology.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXktlGgtrs%3D&md5=849ad7f3d54edb1482b75bffb6411705CAS | 19276618PubMed |

Kerner, J., and Hoppel, C. (2000). Fatty acid import into mitochondria. Biochim. Biophys. Acta 1486, 1–17.
| 1:CAS:528:DC%2BD3cXlt1Snt74%3D&md5=b5da1f579f77af8ad4ca0a6ada71a383CAS | 10856709PubMed |

Kikuchi, K., Nagai, T., Ding, J., Yamauchi, N., Noguchi, J., and Izaike, Y. (1999). Cytoplasmic maturation for activation of pig follicular oocytes cultured and arrested at metaphase I. J. Reprod. Fertil. 116, 143–156.
Cytoplasmic maturation for activation of pig follicular oocytes cultured and arrested at metaphase I.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXjvVemsbk%3D&md5=b00f7ba427b19150f3b996e502f33c46CAS | 10505065PubMed |

Kikuchi, K., Onishi, A., Kashiwazaki, N., Iwamoto, M., Noguchi, J., Kaneko, H., Akita, T., and Nagai, T. (2002a). Successful piglet production after transfer of blastocysts produced by a modified in vitro system. Biol. Reprod. 66, 1033–1041.
Successful piglet production after transfer of blastocysts produced by a modified in vitro system.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XitlClur8%3D&md5=269b24fb5177410d3dbd3914a3a1cb9bCAS | 11906923PubMed |

Kikuchi, K., Ekwall, H., Tienthai, P., Kawai, Y., Noguchi, J., Kaneko, H., and Rodriguez-Martinez, H. (2002b). Morphological features of lipid droplet transition during porcine oocyte fertilisation and early embryonic development to blastocyst in vivo and in vitro. Zygote 10, 355–366.
Morphological features of lipid droplet transition during porcine oocyte fertilisation and early embryonic development to blastocyst in vivo and in vitro.Crossref | GoogleScholarGoogle Scholar | 12463532PubMed |

Krisher, R. L., Brad, A. M., Herrick, J. R., Sparman, M. L., and Swain, J. E. (2007). A comparative analysis of metabolism and viability in porcine oocytes during in vitro maturation. Anim. Reprod. Sci. 98, 72–96.
A comparative analysis of metabolism and viability in porcine oocytes during in vitro maturation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXhs1Sksrw%3D&md5=c68a63c9df9130302b75ca30504e4298CAS | 17110061PubMed |

McEvoy, T. G., Coull, G. D., Broadbent, P. J., Hutchinson, J. S., and Speake, B. K. (2000). Fatty acid composition of lipids in immature cattle, pig and sheep oocytes with intact zona pellucida. J. Reprod. Fertil. 118, 163–170.
| 1:CAS:528:DC%2BD3cXpsl2msA%3D%3D&md5=4996e9d6cefc07c983d3541dad5ac072CAS | 10793638PubMed |

Men, H., Agca, Y., Riley, L. K., and Critser, J. K. (2006). Improved survival of vitrified porcine embryos after partial delipation through chemically stimulated lipolysis and inhibition of apoptosis. Theriogenology 66, 2008–2016.
Improved survival of vitrified porcine embryos after partial delipation through chemically stimulated lipolysis and inhibition of apoptosis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XhtVGjtL3K&md5=d2b044b954a7b4428ea309ccf9b2f9a8CAS | 16870242PubMed |

Nasr-Esfahani, M. H., Aitken, J. R., and Johnson, M. H. (1990). Hydrogen peroxide levels in mouse oocytes and early cleavage stage embryos developed in vitro or in vivo. Development 109, 501–507.
| 1:CAS:528:DyaK3cXlsV2nsbo%3D&md5=cf2046f101d7a6368f46b999f1cee07aCAS | 2401209PubMed |

Petters, R. M., and Wells, K. D. (1993). Culture of pig embryos. J. Reprod. Fertil. Suppl. 48, 61–73.
| 1:STN:280:DyaK2c7psVCktQ%3D%3D&md5=1693e6b3d03fbbac8f6082018c0b977eCAS | 8145215PubMed |

Pillich, R. T., Scarsella, G., and Risuleo, G. (2005). Reduction of apoptosis through the mitochondrial pathway by the administration of acetyl-L-carnitine to mouse fibroblasts in culture. Exp. Cell Res. 306, 1–8.
Reduction of apoptosis through the mitochondrial pathway by the administration of acetyl-L-carnitine to mouse fibroblasts in culture.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXjvVOht7o%3D&md5=5deb2b283358c71616344f221f613eebCAS | 15878327PubMed |

Somfai, T., Kikuchi, K., Medvedev, S., Onishi, A., Iwamoto, M., Fuchimoto, D., Ozawa, M., Noguchi, J., Kaneko, H., Ohnuma, K., Sato, E., and Nagai, T. (2005). Development to the blastocyst stage of immature pig oocytes arrested before the metaphase-II stage and fertilized in vitro. Anim. Reprod. Sci. 90, 307–328.
Development to the blastocyst stage of immature pig oocytes arrested before the metaphase-II stage and fertilized in vitro.Crossref | GoogleScholarGoogle Scholar | 15899560PubMed |

Stojkovic, M., Machado, S. A., Stojkovic, P., Zakhartchenko, V., Hutzler, P., Gonçalves, P. B., and Wolf, E. (2001). Mitochondrial distribution and adenosine triphosphate content of bovine oocytes before and after in vitro maturation: correlation with morphological criteria and developmental capacity after in vitro fertilization and culture. Biol. Reprod. 64, 904–909.
Mitochondrial distribution and adenosine triphosphate content of bovine oocytes before and after in vitro maturation: correlation with morphological criteria and developmental capacity after in vitro fertilization and culture.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXhsVKjtrk%3D&md5=1c33fa33a77fabe3ffd8504ad8c8ffc9CAS | 11207207PubMed |

Sturmey, R. G., and Leese, H. J. (2003). Energy metabolism in pig oocytes and early embryos. Reproduction 126, 197–204.
Energy metabolism in pig oocytes and early embryos.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXntFyjtr4%3D&md5=909ebac0d2348d75ceaeafecb234253bCAS | 12887276PubMed |

Sturmey, R. G., O’Toole, P. J., and Leese, H. J. (2006). Fuorescence resonance energy transfer analysis of mitochondrial:lipid association in the porcine oocyte. Reproduction 132, 829–837.
Fuorescence resonance energy transfer analysis of mitochondrial:lipid association in the porcine oocyte.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXmsF2isw%3D%3D&md5=c17b2984a3f8cdf72a8c849e47788d4aCAS | 17127743PubMed |

Sturmey, R. G., Reis, A., Leese, H. J., and McEvoy, T. G. (2009). Role of fatty acids in energy provision during oocyte maturation and early embryo development. Reprod. Domest. Anim. 44, 50–58.
Role of fatty acids in energy provision during oocyte maturation and early embryo development.Crossref | GoogleScholarGoogle Scholar | 19660080PubMed |

Sun, Q. Y., Wu, G. M., Lai, L., Park, K. W., Cabot, R., Cheong, H. T., Day, B. N., Prather, R. S., and Schatten, H. (2001). Translocation of active mitochondria during pig oocyte maturation, fertilization and early embryo development in vitro. Reproduction 122, 155–163.
Translocation of active mitochondria during pig oocyte maturation, fertilization and early embryo development in vitro.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXlsVGis7g%3D&md5=f3bd0daf3ae7c954ed66960d68b01ed8CAS | 11425340PubMed |

Suzuki, K., Asano, A., Eriksson, B., Niwa, K., Nagai, T., and Rodriguez-Martinez, H. (2002). Capacitation status and in vitro fertility of boar spermatozoa: effects of seminal plasma, cumulus-oocyte-complexes-conditioned medium and hyaluronan. Int. J. Androl. 25, 84–93.
Capacitation status and in vitro fertility of boar spermatozoa: effects of seminal plasma, cumulus-oocyte-complexes-conditioned medium and hyaluronan.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XjtFSitb8%3D&md5=86340bf29598b0eb4fe4eb34c83393a6CAS | 11903657PubMed |

Tatemoto, H., Ootaki, K., Shigeta, K., and Muto, N. (2001). Enhancement of developmental competence after in vitro fertilization of porcine oocytes by treatment with ascorbic acid 2-O-alpha-glucoside during in vitro maturation. Biol. Reprod. 65, 1800–1806.
Enhancement of developmental competence after in vitro fertilization of porcine oocytes by treatment with ascorbic acid 2-O-alpha-glucoside during in vitro maturation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXos1Kntr0%3D&md5=38f923de2606de7680e725e414860bdcCAS | 11717144PubMed |

Vanella, A., Russo, A., Acquaviva, R., Campisi, A., Di Giacomo, C., Sorrenti, V., and Barcellona, M. L. (2000). L-propionyl-carnitine as superoxide scavenger, antioxidant, and DNA cleavage protector. Cell Biol. Toxicol. 16, 99–104.
L-propionyl-carnitine as superoxide scavenger, antioxidant, and DNA cleavage protector.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXlvVWntro%3D&md5=f0c4462fab76774c51d6dd91575ea88fCAS | 10917565PubMed |

Yamada, T., Imai, H., and Yamada, M. (2006). Beneficial effects of acetyl-L-carnitine treatment during IVM on post-fertilization development of bovine oocytes in vitro. Reprod. Fertil. Dev. 18, 280–281.
Beneficial effects of acetyl-L-carnitine treatment during IVM on post-fertilization development of bovine oocytes in vitro.Crossref | GoogleScholarGoogle Scholar |

Ye, J., Li, J., Yu, Y., Wei, Q., Deng, W., and Yu, L. (2010). L-carnitine attenuates oxidant injury in HK-2 cells via ROS-mitochondria pathway. Regul. Pept. 161, 58–66.
L-carnitine attenuates oxidant injury in HK-2 cells via ROS-mitochondria pathway.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXjvFKisb4%3D&md5=9ccd577ceb21634f36121b9c6f2cae2cCAS | 20093144PubMed |

Yu, Y., Dumollard, R., Rossbach, A., Lai, F. A., and Swann, K. (2010). Redistribution of mitochondria leads to bursts of ATP production during spontaneous mouse oocyte maturation. J. Cell. Physiol. 224, 672–680.
Redistribution of mitochondria leads to bursts of ATP production during spontaneous mouse oocyte maturation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXosVWntLo%3D&md5=72478d66035798e59f7b77cef72d83afCAS | 20578238PubMed |