Monteiro F M, Ferreira M M G, Potiens J R, Eberhardt B G, Trinca L A, Barros C M
Department of Pharmacology, Institute of Bioscience, UNESP, Botucatu, Sao Paulo, Brazil.
Reprod Domest Anim. 2010 Oct;45(5):860-4. doi: 10.1111/j.1439-0531.2009.01399.x.
There are indications in the literature that delaying the period between ovarian superestimulation and ovum pick up (OPU) would induce follicles to a condition of initial atresia, which could be beneficial to oocyte development. In this work, we compared three protocols for OPU and in vitro production (IVP) of embryos, in Nellore cattle. Nellore cows (n = 18) were randomly allocated in three groups: Group 1 (OPU), Group 2 [Follicle stimulating hormone (FSH) and OPU] and Group 3 (FSH deprivation and OPU). Three OPUs were performed, and the animals were switched to a different group each time (crossover), in such a way that at the end of the experiment all cows received the 3 protocols. At random stage of the oestrous cycle (D-2), all follicles ≥ 6 mm were aspirated to induce a new follicular wave 2 days afterwards (D0). In Group 1, OPU was performed on D2 and oocytes were processed to IVP. In Group 2, starting on D0, cows were superstimulated (FSH, Folltropin(®), 30 mg administered daily, i.m., during three consecutive days, total dose = 180 mg), and 6 h after the last FSH dose, they received exogenous luteinizing hormone (LH) (12.5 mg, i.m., Lutropin(®), D3). The OPU was performed 6 h after LH administration, i.e. 12 h after the last dose of FSH. Animals in Group 3 received the same treatment as those in Group 2, except that LH was administered 42 h after the last dose of FSH, and OPU occurred 6 h later. Therefore, in this group, follicles were deprived of FSH at 48 h. Both cleavage and blastocyst rates were similar (p > 0.05, anova) among oocytes from Groups 1, 2 and 3, respectively: 77.4% (144/185) and 42.70% (79/185); 75.54% (105/139) and 31.65% (44/139); 63.52% (101/159) and 33.33% (53/159). However, hatched blastocyst rate was higher (p < 0.01) in Group 1 (30.27%, 56/185) when compared with Group 2 (11.51%, 16/139) or 3 (15.72%, 25/159). It is concluded that, contrary to previous work on European breeds (Bos taurus), ovarian superstimulation associated with deprivation of FSH and OPU (Group 3) did not increase IVP of Nellore embryos (Bos indicus). On the contrary, the highest hatched blastocyst rates were observed in oocytes from non-superstimulated cows.
文献中有迹象表明，推迟卵巢超刺激与采卵（OPU）之间的时间间隔会使卵泡进入初始闭锁状态，这可能有利于卵母细胞的发育。在本研究中，我们比较了内洛尔牛的三种OPU及胚胎体外生产（IVP）方案。将18头内洛尔母牛随机分为三组：第1组（OPU组）、第2组[促卵泡素（FSH）和OPU组]和第3组（FSH剥夺和OPU组）。进行了三次OPU，每次将动物切换到不同的组（交叉分组），使得在实验结束时所有母牛都接受了这三种方案。在发情周期的随机阶段（第-2天），抽吸所有直径≥6mm的卵泡，以在两天后（第0天）诱导新的卵泡波。在第1组中，于第2天进行OPU，并将卵母细胞用于IVP。在第2组中，从第0天开始，母牛接受超刺激（FSH，Folltropin®，每天30mg，肌肉注射，连续三天，总剂量=180mg），在最后一剂FSH注射后6小时，它们接受外源性促黄体生成素（LH）（12.5mg，肌肉注射，Lutropin®，第3天）。在LH注射后6小时进行OPU，即最后一剂FSH注射后12小时。第3组的动物接受与第2组相同的处理，不同之处在于LH在最后一剂FSH注射后42小时给药，OPU在6小时后进行。因此，在该组中，卵泡在48小时时被剥夺了FSH。第1、2和3组的卵母细胞的分裂率和囊胚率分别相似（p>0.05，方差分析）：77.4%（144/185）和42.70%（79/185）；75.54%（105/139）和31.65%（44/139）；63.52%（101/159）和33.33%（53/159）。然而，与第2组（11.51%，16/139）或第3组（15.72%，25/159）相比，第1组的孵化囊胚率更高（p<0.01）（30.27%，56/185）。得出的结论是，与先前关于欧洲品种（Bos taurus）的研究相反，与FSH剥夺和OPU相关的卵巢超刺激（第3组）并未增加内洛尔胚胎（Bos indicus）的IVP。相反，在未接受超刺激的母牛的卵母细胞中观察到最高的孵化囊胚率。
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