Laboratorio de Producción Equina, Facultad de Agronomía y Veterinaria, Universidad Nacional de Río Cuarto, Río Cuarto, Córdoba, Argentina; Consejo Nacional de Investigaciones Científicas y Técnicas, Ciudad Autónoma de Buenos Aires, Buenos Aires, Argentina.
Veterinary Reproduction Group, Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, University of Cordoba, 14014, Cordoba, Spain.
Theriogenology. 2023 Jan 1;195:199-208. doi: 10.1016/j.theriogenology.2022.10.030. Epub 2022 Nov 1.
Equus members exhibit very divergent karyotype, genetic plasticity, and significant differences in their reproductive physiology. Despite the fact that somatic cell nuclear transfer and intracytoplasmic sperm injection (ICSI) has gained relevance in the last few years in horses, few reports have been published exploring ovum pick up (OPU) and in vitro maturation (IVM) of cumulus-oocyte complexes (COCs) in donkeys. Yet, some donkey species and breeds are considered endangered, and these assisted-reproductive technologies could help to preserve the genetic of valuable individuals. In this study, we tested the hypothesis that supplementation with jenny preovulatory follicular fluid (PFF) during IVM could improve oocyte developmental competence in the donkey. For this, in vitro nuclear maturation rates, cumulus cell expansion, and embryo development after ICSI of donkey COCs matured in culture media supplemented with fetal bovine serum (FBS) or donkey PFF, with a known metabolomic profile, were assessed. Time-lapse imagining was performed after ICSI of horse and donkey oocytes. Eight OPU sessions were done in five jennies with an average recovery rate of 69.2% (n = 45 COCs). Although lower cumulus cells expansion was observed in oocytes of PFF group (P = 0.0010), no significant differences were described in nuclear maturation rates and preimplantation embryo development between groups. Donkey ICSI embryos showed similar morphokinetics to horse ICSI embryos. Our study shows that supplementing IVM media with FBS or donkey PFF supports nuclear maturation and early preimplantation embryo development after ICSI in donkeys. To our knowledge, the present study is the first report of ICSI, time-lapse imaging and in vitro blastocyst production in donkey.
马属动物的核型表现出很大的差异，遗传可塑性强，生殖生理学也有很大的差异。尽管近年来体细胞核移植和胞质内精子注射（ICSI）在马中得到了重视，但很少有报道探讨驴卵母细胞的卵母细胞采集（OPU）和体外成熟（IVM）。然而，一些驴种和品种被认为处于濒危状态，这些辅助生殖技术可以帮助保存有价值个体的遗传资源。在这项研究中，我们检验了这样一个假设，即在 IVM 期间补充母马发情前卵泡液（PFF）可以提高驴卵母细胞的发育能力。为此，我们评估了在添加有胎牛血清（FBS）或已知代谢组学特征的驴 PFF 的培养基中成熟的驴 COC 的体外核成熟率、卵丘细胞扩张和 ICSI 后胚胎发育。在 ICSI 后进行了马和驴卵母细胞的延时成像。在五只母马中进行了 8 次 OPU 手术，平均回收率为 69.2%（n=45 COCs）。尽管 PFF 组的卵丘细胞扩张较小（P=0.0010），但两组间核成熟率和植入前胚胎发育无显著差异。驴 ICSI 胚胎与马 ICSI 胚胎表现出相似的形态动力学特征。我们的研究表明，在 IVM 培养基中添加 FBS 或驴 PFF 支持核成熟和 ICSI 后的早期植入前胚胎发育。据我们所知，本研究首次报道了驴的 ICSI、延时成像和体外囊胚生产。
