Rodríguez María Belén, Gambini Andrés, Clérico Gabriel, Ynsaurralde-Rivolta Amada Eugenia, Briski Olinda, Largel Hernan, Sansinena Marina, Salamone Daniel F
Laboratorio de Biotecnología Animal, Facultad de Agronomía, Universidad de Buenos Aires, Av. San Martin 4453, Agronomía, C1417DSE CABA, Buenos Aires, Argentina; and Consejo Nacional de Investigaciones Científicas y Técnicas, (CONICET) Godoy Cruz 2290, C1425FQB CABA, Argentina.
Consejo Nacional de Investigaciones Científicas y Técnicas, (CONICET) Godoy Cruz 2290, C1425FQB CABA, Argentina; and Laboratorio de Biotecnología y Reproducción Animal, Facultad de Ingeniería y Ciencias Agrarias, Pontificia Universidad Católica Argentina, Av. Alicia Moreau de Justo 1300, CABA, Buenos Aires, Argentina.
Reprod Fertil Dev. 2019 Jan;31(12):1805-1811. doi: 10.1071/RD19248.
Assisted reproduction techniques (ARTs) have become widespread in the equine breeding industry. In particular, the combination of oocyte recovery from live mares followed by IVM and intracytoplasmic sperm injection (ICSI) has increased markedly among the ARTs used with valuable or low-fertility animals. There is currently no consensus among research groups regarding the optimal oocyte maturation period to produce high-quality embryos. In this study, we report the maturation dynamics of equine oocytes at different time points, from 20 to 40h (Experiment 1). In addition, in Experiment 2, equine ICSI blastocysts were produced from oocytes that exhibited early (up to 24h) or late (28-30h) extrusion of the first polar body (PB). Blastocyst rates and diameter were recorded and embryo quality was assessed by analysing the number of apoptotic cells and Yes-associated protein 1 (YAP1) expression. By 20h of IVM, 42% of oocytes were mature, and the remaining oocytes matured within the next 17h of IVM. Although no differences were found in cell apoptosis or the number of YAP1-positive cells between groups exhibiting early and late PB extrusion, embryos from the early group (Group I) exhibited an improved total cell number and blastocyst rate compared to embryos from the late group (Group II) (18.60% vs 10.17% respectively).
辅助生殖技术（ARTs）在马的繁殖行业中已广泛应用。特别是，从活体母马采集卵母细胞，随后进行体外成熟（IVM）和卵胞浆内单精子注射（ICSI）的联合技术，在用于珍贵或低生育力动物的ARTs中显著增加。目前，各研究小组对于产生高质量胚胎的最佳卵母细胞成熟期尚未达成共识。在本研究中，我们报告了马卵母细胞在20至40小时不同时间点的成熟动态（实验1）。此外，在实验2中，从第一极体（PB）排出早（最长24小时）或晚（28 - 30小时）的卵母细胞中产生马ICSI囊胚。记录囊胚率和直径，并通过分析凋亡细胞数量和Yes相关蛋白1（YAP1）表达来评估胚胎质量。体外成熟20小时时，42%的卵母细胞成熟，其余卵母细胞在接下来的17小时体外成熟过程中成熟。尽管在PB排出早和晚的两组之间，细胞凋亡或YAP1阳性细胞数量没有差异，但早期组（I组）的胚胎与晚期组（II组）的胚胎相比，总细胞数和囊胚率有所提高（分别为18.60%和10.17%）。
