Crispo M, Meikle M N, Schlapp G, Menchaca A
Unidad de Animales Transgénicos y de Experimentación, Institut Pasteur de Montevideo, Mataojo 2020, Montevideo, Uruguay.
Unidad de Animales Transgénicos y de Experimentación, Institut Pasteur de Montevideo, Mataojo 2020, Montevideo, Uruguay.
Theriogenology. 2021 Apr 1;164:31-35. doi: 10.1016/j.theriogenology.2021.01.012. Epub 2021 Jan 21.
Pharmacological ovarian control required for the implementation of artificial insemination and embryo-related technologies usually includes the use of eCG, naturally produced in pregnant mares. In this study, we report the superovulatory response and embryo development in mice obtained with a new glycoprotein with eCG-like activity (reCG) produced by recombinant DNA technology. A total of 150 females from three different mouse strains (C57BL/6J, BALB/cJ and B6D2F1/J) were subjected to a superstimulatory protocol consisting of 5 IU of natural eCG (n = 50), 5 IU of reCG (n = 50), or received a placebo (no-eCG, n = 50) by intraperitoneal route, followed by 5 IU of human chorionic gonadotropin 48 h later. Overall, no significant differences were observed in the total number of ova/zygotes (33.6 ± 2.4 vs 28.7 ± 2.6; P = NS) and viable ova/zygotes (31.5 ± 2.4 vs 25.8 ± 2.5; P = NS) collected per female among eCG and reCG treated females, respectively, which were greater (P < 0.05) than those obtained in no-eCG treated females (6.9 ± 0.7 and 5.9 ± 0.7, respectively). Zygotes derived from the three experimental groups (n = 2914) were subjected to in vitro culture until hatching 4.5 days post coitum (dpc). Regardless of the mouse strain, no differences were observed among eCG and reCG treated females for overall cleavage rate 1.5 dpc (58.5% vs 60.5%), development rate 3.5 dpc (47.2% vs 48.9%) and hatching rate 4.5 dpc (49.5% vs 54.5) (P = NS). Control females from no-eCG treated group showed lower cleavage and development rates (36.4% and 29.7%, respectively; P < 0.05). In conclusion, this study reports for the first time comparable superovulatory response and embryo development between recombinant and natural eCG treatment, which has important implications for reproductive technologies in several species.
实施人工授精和胚胎相关技术所需的药理学卵巢控制通常包括使用孕马自然产生的eCG。在本研究中,我们报告了用重组DNA技术生产的具有eCG样活性的新型糖蛋白(reCG)处理的小鼠的超排卵反应和胚胎发育情况。来自三种不同小鼠品系(C57BL/6J、BALB/cJ和B6D2F1/J)的总共150只雌性小鼠接受了超刺激方案,通过腹腔途径注射5 IU天然eCG(n = 50)、5 IU reCG(n = 50)或接受安慰剂(无eCG,n = 50),48小时后再注射5 IU人绒毛膜促性腺激素。总体而言,eCG和reCG处理的雌性小鼠每只收集的卵子/合子总数(33.6 ± 2.4对28.7 ± 2.6;P = 无显著性差异)和存活卵子/合子数(31.5 ± 2.4对25.8 ± 2.5;P = 无显著性差异)之间未观察到显著差异,这两组均高于未接受eCG处理的雌性小鼠(分别为6.9 ± 0.7和5.9 ± 0.7,P < 0.05)。来自三个实验组(n = 2914)的合子在体外培养至交配后4.5天(dpc)孵化。无论小鼠品系如何,eCG和reCG处理的雌性小鼠在1.5 dpc的总体卵裂率(58.5%对60.5%)、3.5 dpc的发育率(47.2%对48.9%)和4.5 dpc的孵化率(49.5%对54.5)方面均未观察到差异(P = 无显著性差异)。未接受eCG处理组的对照雌性小鼠显示出较低的卵裂率和发育率(分别为36.4%和29.7%;P < 0.05)。总之,本研究首次报告了重组eCG和天然eCG处理之间可比的超排卵反应和胚胎发育情况,这对多个物种的生殖技术具有重要意义。