Moura Rodrigo, Fernandes Carlos Antonio, Siqueira Luiz Gustavo, Figueiredo Ricardo Alamino, Martins Carlos Frederico, Peixer Maurício Antonio, Xavier Marcelo Cunha, Viana Joao Henrique
Universidade de Brasília, Brasília, DF, 70910-900, Brazil.
Universidade Edson Antônio Velano, Alfenas, MG, 37130-000, Brazil.
Theriogenology. 2025 Mar 1;234:110-116. doi: 10.1016/j.theriogenology.2024.12.011. Epub 2024 Dec 11.
The aim of this study was to evaluate the use of a long-acting recombinant human FSH (rhFSH, corifollitropin-alpha) to induce ovarian stimulation in Nelore breed (Bos indicus) calves and prepubertal heifers prior to ovum pick-up (OPU) for in vitro embryo production (IVEP). In Experiment 1, a dose-response trial was performed to determine the optimal dose of rhFSH, which was determined to be 10 μg. In Experiment 2, 6-7 mo old calves were randomly allocated to receive rhFSH either via sc (n = 5) or im (n = 5). Ovarian follicular development was monitored daily by transrectal ultrasonography for five days. There was no effect of route (P = 0.1348) nor route × time interaction (P = 0.8336) on follicle development. In Experiment 3, 7-8 mo old calves (n = 90) were randomly allocated into 5 groups: 1) Control: no ovarian stimulation; 2) rhFSH-96: 10 μg rhFSH sc followed by OPU 96h later; 3) rhFSH-120: 10 μg rhFSH sc and OPU 120h later; 4) eCG-96: 300 IU eCG im and OPU 96h later; and 5) eCG-120: 300 IU eCG im and OPU 120h later. Non-rhFSH treated Nelore mature cows (n = 10) were used as reference-controls for IVEP outcomes. Treatment with rhFSH increased the proportion of grade I cumulus-oocyte complexes (COC) collected by OPU at either timepoints (96 or 120h) compared with eCG or controls (P < 0.0001). Blastocyst rate for rhFSH-120 calves was similar to mature cows (P > 0.05). Treatment with rhFSH, however, increased the proportion of expanded COC and decreased the proportion of viable COC (P < 0.0001) compared with eCG and controls. In Experiment 4, yearling heifers (n = 60) were treated or not (control group) with 10 μg rhFSH sc and OPU was performed 72 or 96h later. Heifers treated with rhFSH had a greater proportion of grade I COC (P = 0.0188) and blastocyst rate (P < 0.0098) than controls, regardless of the interval used. These groups, however, yielded a lesser number of viable COC (P = 0.0264), resulting in a similar (P = 0.5869) number of embryos produced by donor per OPU compared with controls. Pregnancy rate after embryo transfer was also similar between controls and rhFSH groups (19.3 vs 25.0 %, P = 0.4142). In summary, treatment with a single sc injection of corifollitropin-alpha was effective to promote ovarian stimulation in calves prior to OPU. The potential benefits of stimulatory protocols using rhFSH, however, have been overshadowed by a decrease in the total number of viable COC recovered per donor, thus failing to increase the number of embryos produced per OPU.
本研究的目的是评估使用长效重组人促卵泡激素(rhFSH,阿那曲唑)在体外胚胎生产(IVEP)的采卵(OPU)前诱导瘤牛(印度牛)犊牛和青春期前小母牛的卵巢刺激。在实验1中,进行了剂量反应试验以确定rhFSH的最佳剂量,确定为10μg。在实验2中,将6-7月龄的犊牛随机分配接受rhFSH,通过皮下注射(n = 5)或肌肉注射(n = 5)。通过经直肠超声检查每天监测卵巢卵泡发育,持续5天。在卵泡发育方面,途径(P = 0.1348)和途径×时间交互作用(P = 0.8336)均无影响。在实验3中,将7-8月龄的犊牛(n = 90)随机分为5组:1)对照组:不进行卵巢刺激;2)rhFSH-96:皮下注射10μg rhFSH,96小时后进行OPU;3)rhFSH-120:皮下注射10μg rhFSH,120小时后进行OPU;4)eCG-96:肌肉注射300IU eCG,96小时后进行OPU;5)eCG-120:肌肉注射300IU eCG,120小时后进行OPU。未用rhFSH处理的瘤牛成年母牛(n = 10)用作IVEP结果的参考对照。与eCG或对照组相比,rhFSH处理增加了在任何时间点(96或120小时)通过OPU收集的I级卵丘-卵母细胞复合体(COC)的比例(P <0.0001)。rhFSH-120组犊牛的囊胚率与成年母牛相似(P> 0.05)。然而,与eCG和对照组相比,rhFSH处理增加了扩张的COC的比例并降低了活COC的比例(P <0.0001)。在实验4中,对一岁小母牛(n = 60)进行或不进行(对照组)10μg rhFSH皮下注射,并在72或96小时后进行OPU。无论使用何种间隔,用rhFSH处理的小母牛的I级COC比例(P = 0.0188)和囊胚率(P <0.0098)均高于对照组。然而,这些组产生的活COC数量较少(P = 0.0264),导致与对照组相比,每个OPU供体产生的胚胎数量相似(P = 0.5869)。胚胎移植后的妊娠率在对照组和rhFSH组之间也相似(19.3对25.0%,P = 0.4142)。总之,单次皮下注射阿那曲唑治疗可有效促进犊牛在OPU前的卵巢刺激。然而,使用rhFSH的刺激方案的潜在益处因每个供体回收的活COC总数减少而被掩盖,因此未能增加每个OPU产生的胚胎数量。