Follicular growth-stimulated cows provide favorable oocytes for producing cloned embryos.
We examined the influence of recipient oocytes on in vitro development, oxygen consumption, and gene expression in the resulting cloned bovine embryos. Oocytes derived from slaughterhouse ovaries and ovum pickup (OPU)-derived oocytes were used as recipient cytoplasts for the production of cloned embryos. A series of OPU sessions was conducted on Holstein cows without follicular growth treatment (FGT). In the same cows, we then performed dominant follicle ablation and subsequently administered follicle-stimulating hormone and prostaglandin F(2α) with controlled internal drug release device before a second series of OPU. Cumulus cells collected from single Holstein cows were used as donor cells. After measurement of oxygen consumption at the blastocyst stage with modified scanning electrochemical microscopy, analysis of 10 genes (CDX2, IFN-tau, PLAC8, OCT4, SOX2, NANOG, ATP5A1, GLUT1, AKR1B1, and IGF2R) was performed with real-time RT-PCR. Rates of fusion, cleavage, and blastocyst formation were not different among the treatment groups. Levels of oxygen consumption in cloned blastocysts derived from slaughterhouse ovaries or OPU without FGT were significantly lower than in blastocysts derived from artificial insemination (AI). However, oxygen consumption was increased in cloned blastocysts derived from OPU with FGT, depending on the individual oocyte donor. Furthermore, gene expression of IFN-tau and OCT4 in cloned blastocysts derived from OPU with FGT was similar to that in AI-derived blastocysts, whereas expression of those genes in cloned blastocysts derived from slaughterhouse ovaries or OPU without FGT was significantly different from that in AI-derived blastocysts. Thus, recipient oocytes collected by OPU in combination with manipulation of follicular growth in donor cows are suitable for producing cloned embryos.
National Livestock Breeding Center, Nishigo, Fukushima, Japan.
Satoshi
S
Sugimura
Shuji
S
Kobayashi
Yutaka
Y
Hashiyada
Masaki
M
Ohtake
Masahiro
M
Kaneda
Tadayuki
T
Yamanouchi
Hideo
H
Matsuda
Yoshio
Y
Aikawa
Shinya
S
Watanabe
Takashi
T
Nagai
Eiji
E
Kobayashi
Kazuyuki
K
Konishi
Kei
K
Imai
D000818:Animals; D002417:Cattle; D019976:Cloning, Organism / Q000379:methods*; D015237:Dinoprost / Q000494:pharmacology; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D066298:In Vitro Techniques; D007370:Interferon Type I / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D009865:Oocytes / Q000166:cytology* / Q000502:physiology*; D006080:Ovarian Follicle / Q000187:drug effects / Q000254:growth & development*; D010101:Oxygen Consumption / Q000502:physiology; D011257:Pregnancy Proteins / Q000378:metabolism
D007370:Interferon Type I; D050814:Octamer Transcription Factor-3; D011257:Pregnancy Proteins; C020947:interferon tau; D005640:Follicle Stimulating Hormone; D015237:Dinoprost