title: Optimizing production of in vivo-matured oocytes from superstimulated Holstein cows for in vitro production of embryos using X-sorted sperm.
abstract: The present study aimed to establish an efficient system for the production of female embryos from dairy cows by in vitro fertilization (IVF) using X-sorted sperm and in vivo-matured oocytes collected by ovum pick up (OPU). Nonlactating Holstein cows (n = 36) were administered a controlled intravaginal progesterone-releasing (controlled internal drug release) device (d 0), underwent dominant follicle ablation (DFA) or ovulation by administration of 100 μg of GnRH on d 5, and were superstimulated with FSH and PGF2α, following standard procedures. Controlled internal drug release devices were removed on the evening of d 8 or on the morning of d 9, depending on the experiment. For LH surge induction, 200 μg of GnRH was administered on the morning of d 10 (0 h). In experiment 1, the peak (48.1%) of ovulating follicles was detected at 29 to 32 h after GnRH injection (0 h), and the range in the timing of the initiation of ovulation was less by timing from GnRH administration (30.0 ± 2.8h) rather than by timing the onset of estrus (32.7 ± 4.7h). Only 0.9% of total ovulated follicles were recorded before 26 h after GnRH injection. Therefore, OPU was carried out at 26 h and IVF occurred at 30 h after GnRH in experiments 2 and 3. In experiment 2, 83.3 ± 10.8% of oocytes with expanded cumulus cells had extruded the first polar body at 30 h after GnRH injection. The aim of experiment 3 was to compare the effect of either DFA or GnRH-induced LH surge before superstimulation on the efficiency of embryo production by IVF following superstimulation. Progesterone concentrations from d 10 to 12 in the DFA group were lower than those in the GnRH group. A greater proportion of recovered oocytes with expanded cumulus cells from ≥ 8-mm follicles was observed in the DFA group than in the GnRH group (95.9 and 77.4%, respectively). Blastocyst rates in the DFA and GnRH groups (58.0 and 52.8%, respectively) did not differ from those of oocytes collected from nonstimulated OPU and matured in vitro (49.9%). However, the proportion of high-quality blastocysts was higher in the DFA group compared with the GnRH group (54.9 vs. 21.5%). Our results demonstrate that high rates of good-quality blastocysts can be produced by IVF with X-sorted frozen sperm using in vivo-matured oocytes collected by OPU from cows after DFA and superstimulation combined with ovulation induction.
authors.affiliation: Reproductive Biology and Technology Division, National Livestock Breeding Center, 1 Odakurahara, Nishigo, Fukushima 961-8511, Japan. Electronic address: matoba@affrc.go.jp.
authors.forename: S
authors.initials: S
authors.lastname: Matoba
authors.affiliation: Reproductive Biology and Technology Division, National Livestock Breeding Center, 1 Odakurahara, Nishigo, Fukushima 961-8511, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Yoshioka
authors.affiliation: Reproductive Biology and Technology Division, National Livestock Breeding Center, 1 Odakurahara, Nishigo, Fukushima 961-8511, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Matsuda
authors.affiliation: Reproductive Biology and Technology Division, National Livestock Breeding Center, 1 Odakurahara, Nishigo, Fukushima 961-8511, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Sugimura
authors.affiliation: Reproductive Biology and Technology Division, National Livestock Breeding Center, 1 Odakurahara, Nishigo, Fukushima 961-8511, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Aikawa
authors.affiliation: Reproductive Biology and Technology Division, National Livestock Breeding Center, 1 Odakurahara, Nishigo, Fukushima 961-8511, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Ohtake
authors.affiliation: Reproductive Biology and Technology Division, National Livestock Breeding Center, 1 Odakurahara, Nishigo, Fukushima 961-8511, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Hashiyada
authors.affiliation: Niigata Agricultural Research Institute Livestock Research Center, 178 Tanahire, Sanjo, Niigata 955-0143, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Seta
authors.affiliation: Niigata Agricultural Research Institute Livestock Research Center, 178 Tanahire, Sanjo, Niigata 955-0143, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Nakagawa
authors.affiliation: School of Agriculture and Food Science, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: P
authors.initials: P
authors.lastname: Lonergan
authors.affiliation: Reproductive Biology and Technology Division, National Livestock Breeding Center, 1 Odakurahara, Nishigo, Fukushima 961-8511, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Imai
mesh_terms: D000818:Animals; D002417:Cattle; D015237:Dinoprost / Q000378:metabolism; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D007987:Gonadotropin-Releasing Hormone / Q000494:pharmacology*; D008297:Male; D054315:Oocyte Retrieval / Q000662:veterinary*; D009865:Oocytes / Q000166:cytology*; D010062:Ovulation Induction / Q000662:veterinary; D011374:Progesterone / Q000378:metabolism; D013094:Spermatozoa / Q000502:physiology
chemical_list: D007987:Gonadotropin-Releasing Hormone; D011374:Progesterone; D015237:Dinoprost
keywords: in vitro fertilization; in vivo-matured oocyte; sex-sorted sperm; superstimulation