da Silva Júlio César Barboza, Ferreira Roberta Machado, Maturana Filho Milton, Naves Julianne de Rezende, Santin Thiago, Pugliesi Guilherme, Madureira Ed Hoffmann
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo (USP), Pirassununga, SP 13635-900, Brazil.
Department of Animal Reproduction, College of Veterinary Medicine and Animal Science, University of São Paulo (USP), Pirassununga, SP 13635-900, Brazil.
Theriogenology. 2017 Mar 1;90:65-73. doi: 10.1016/j.theriogenology.2016.11.016. Epub 2016 Nov 18.
We aimed with the present study to evaluate the effects of FSH treatment (200 mg) split in four or six administrations on ovarian follicle stimulation and in vitro oocyte competence for embryo production in dairy cows with synchronized follicular wave emergence. On random days of the estrous cycle (Day 0), non-lactating Holstein cows received a progesterone (P4)-releasing intravaginal device and 2 mg estradiol benzoate IM. On Day 3, they received 0.530 mg sodium cloprostenol (PGF2α) IM. Control cows (n = 35) received no further treatments, whereas FSH-treated cows received 200 mg FSH split in four (FSH4 group; n = 33) or six (FSH6 group; n = 33) administration regimens. Starting on Day 4, cows in FSH4 group received 200 mg FSH split in four equivalent doses of 50 mg 12 h apart. Cows in FSH6 group received the same total FSH dose split in six equivalent doses of 33.3 mg 12 h apart, but treatments started on Day 3. On Day 7 AM (36 h of "coasting" period for FSH-treated groups), the P4 devices were removed and cows were subjected to ovum pick up (OPU). Viable oocytes were in vitro fertilized using sexed-sorted semen. Although FSH treatment did not (P > 0.1) increase the total number of follicles (Control, 53.2 ± 4.5 vs. FSH-treated, 51.4 ± 3.1), the two hormonal stimulation regimens, FSH4 and FSH6, increased the number of medium follicles (6-10 mm; 5.2 ± 0.5 vs. 18.1 ± 1.4; P < 0.0001) and reduced the number of small follicles (2-5.9 mm; 46.3 ± 5.1 vs. 31.0 ± 2.4 P < 0.0001). Also, FSH treatment or regimen did not increase (P > 0.1) the number of viable oocytes (Control, 12.6 ± 1.26 vs. FSH-treated, 12.70 ± 1.03), recovery rate (Control, 36.5% vs. FSH-treated, 36%) and the number of in vitro produced blastocyst (Control, 4.1 ± 0.52 vs. FSH-treated 4.3 ± 0.5). We concluded that FSH stimulation protocol proposed herein is effective to stimulate the growth of small antral follicle population prior to OPU, but it was ineffective to improve in vitro oocyte competence for embryo production in non-lactating Holstein cows with synchronized follicular wave emergence.
本研究旨在评估将200毫克促卵泡素（FSH）分四次或六次给药，对处于卵泡波同步出现状态的奶牛进行卵巢卵泡刺激以及对体外胚胎生产中卵母细胞能力的影响。在发情周期的随机日期（第0天），非泌乳荷斯坦奶牛接受一个释放孕酮（P4）的阴道内装置以及2毫克苯甲雌二醇肌肉注射。在第3天，它们接受0.530毫克氯前列醇钠（PGF2α）肌肉注射。对照奶牛（n = 35）不再接受进一步处理，而接受FSH处理的奶牛接受200毫克FSH，分为四次给药方案（FSH4组；n = 33）或六次给药方案（FSH6组；n = 33）。从第4天开始，FSH4组的奶牛接受200毫克FSH，分为四个等效剂量，每隔12小时注射50毫克。FSH6组的奶牛接受相同总量的FSH，分为六个等效剂量，每隔12小时注射33.3毫克，但给药从第3天开始。在第7天上午（FSH处理组“延缓期”36小时后），取出P4装置，对奶牛进行采卵（OPU）。使用性别分选精液对有活力的卵母细胞进行体外受精。尽管FSH处理并未（P > 0.1）增加卵泡总数（对照，53.2 ± 4.5 vs. FSH处理组，51.4 ± 3.1），但两种激素刺激方案，FSH4和FSH6，增加了中等卵泡数量（6 - 10毫米；5.2 ± 0.5 vs. 18.1 ± 1.4；P < 0.0001）并减少了小卵泡数量（2 - 5.9毫米；46.3 ± 5.1 vs. 31.0 ± 2.4，P < 0.0001）。此外，FSH处理或给药方案并未增加（P > 0.1）有活力卵母细胞数量（对照，12.6 ± 1.26 vs. FSH处理组，12.70 ± 1.03）、回收率（对照，36.5% vs. FSH处理组，36%）以及体外生产的囊胚数量（对照，4.1 ± 0.52 vs. FSH处理组4.3 ± 0.5）。我们得出结论，本文提出的FSH刺激方案在采卵前刺激小腔卵泡群体生长是有效的，但在提高处于卵泡波同步出现状态的非泌乳荷斯坦奶牛体外胚胎生产中卵母细胞能力方面是无效的。
