Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Universidad de La Frontera, Temuco, Chile.
Department of Agricultural Sciences and Natural Resources, Faculty of Agriculture and Forestry, Universidad de La Frontera, Temuco, Chile.
Reproduction. 2017 Sep;154(3):307-318. doi: 10.1530/REP-17-0270.
The efficiency of intracytoplasmic sperm injection (ICSI) in the bovine is low compared to other species. It is unknown whether defective oocyte activation and/or sperm head decondensation limit the success of this technique in this species. To elucidate where the main obstacle lies, we used homologous and heterologous ICSI and parthenogenetic activation procedures. We also evaluated whether maturation negatively impacted the early stages of activation after ICSI. Here we showed that injected bovine sperm are resistant to nuclear decondensation by bovine oocytes and this is only partly overcome by exogenous activation. Remarkably, when we used heterologous ICSI, -matured mouse eggs were capable of mounting calcium oscillations and displaying normal PN formation following injection of bovine sperm, although -matured mouse oocytes were unable to do so. Together, our data demonstrate that bovine sperm are especially resistant to nuclear decondensation by -matured oocytes and this deficiency cannot be simply overcome by exogenous activation protocols, even by inducing physiological calcium oscillations. Therefore, the inability of a suboptimal ooplasmic environment to induce sperm head decondensation limits the success of ICSI in the bovine. Studies aimed to improve the cytoplasmic milieu of -matured oocytes and to replicate the molecular changes associated with capacitation and acrosome reaction will deepen our understanding of the mechanism of fertilization and improve the success of ICSI in this species.
与其他物种相比,牛的胞质内精子注射(ICSI)效率较低。目前尚不清楚卵母细胞激活缺陷和/或精子头部去浓缩是否限制了该技术在该物种中的成功。为了阐明主要障碍所在,我们使用了同源和异源 ICSI 和孤雌激活程序。我们还评估了成熟是否对 ICSI 后的早期激活阶段产生负面影响。在这里,我们表明,注入的牛精子对牛卵母细胞的核去浓缩具有抗性,而外源性激活仅部分克服了这种抗性。值得注意的是,当我们使用异源 ICSI 时,-成熟的小鼠卵能够在注入牛精子后引发钙振荡,并显示出正常的 PN 形成,尽管 -成熟的小鼠卵母细胞无法做到这一点。总之,我们的数据表明,牛精子对 -成熟卵母细胞的核去浓缩具有特别强的抗性,这种缺陷不能简单地通过外源性激活方案克服,即使通过诱导生理钙振荡也不能克服。因此,不成熟的卵质环境无法诱导精子头部去浓缩,限制了 ICSI 在牛中的成功。旨在改善 -成熟卵母细胞胞质环境并复制与获能和顶体反应相关的分子变化的研究将加深我们对受精机制的理解,并提高该物种 ICSI 的成功率。