Laboratory of Spermatozoa Biology, Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Sao Paulo, Brazil.
Laboratory of in vitro Fertilization, Cloning and Animal Transgenesis, Department of Animal Reproduction, School of Veterinary Medicine and Animal Science, University of Sao Paulo, Sao Paulo, Brazil.
PLoS One. 2018 Jul 10;13(7):e0200273. doi: 10.1371/journal.pone.0200273. eCollection 2018.
Sperm samples used on fertilization strongly influence the in vitro production (IVP) rates. However, sperm traits behind this effect are not stated consistently until now. This study aimed to evaluate the isolated and combined effect of some sperm traits (MB: total motility before Percoll® gradient, MA: total motility after Percoll® gradient, AI: acrosome integrity, MI: membrane integrity, MP: mitochondrial membrane potential, and CR: chromatin resistance) on IVP rates. This is the first study focusing on the isolated effect of distinct traits. For this purpose, the experiment was divided in three steps. In first step, to study behavior of traits sperm samples (n = 63 batches) were analyzed and ranked based on each trait. In second step, samples ranked were selected from target ranks regions and allocated in groups of four to five batches, creating Higher and Lower groups, according to two different approaches. One aimed to form groups that differed to all sperm traits simultaneously (effect of combined traits). The other aimed to form groups that differed only to a single sperm trait while no differences were observed for the remaining traits (effect of each isolated trait). In third step, for each group successfully formed in step 2, sperm samples were individually and prospectively used for IVP. Cleavage, embryo development and blastocyst rates were recorded and compared between Higher and Lower of respective trait groups. Surprisingly, evaluation of isolated effects revealed that lower levels of MB, AI and MP resulted in higher embryo development and blastocyst rates (p<0.05), which was not observed on cleavage rate. We conclude that sperm traits strongly influence embryo development after in vitro fertilization (IVF), affecting the zygote competence to achieve blastocyst stage. Individually, levels of MB, AI or MP could be some of the key traits that may define IVP efficiency on current systems of embryo production.
用于受精的精子样本强烈影响体外生产（IVP）率。然而，直到现在，这种影响背后的精子特征还没有得到一致的说明。本研究旨在评估一些精子特征（MB：Percoll®梯度前的总活力，MA：Percoll®梯度后的总活力，AI：顶体完整性，MI：膜完整性，MP：线粒体膜电位和 CR：染色质抗性）的单独和组合效应对 IVP 率的影响。这是第一项专注于不同特征的单独影响的研究。为此，实验分为三个步骤。在第一步中，为了研究精子样本特征的行为（n = 63 批），根据每个特征对样本进行分析和排序。在第二步中，从目标等级区域中选择排名靠前的样本，并根据两种不同的方法将其分配到四到五个批次的组中，创建高组和低组。一种方法旨在形成同时对所有精子特征有差异的组（综合特征的影响）。另一种方法旨在形成仅对单个精子特征有差异而其余特征没有差异的组（每个单独特征的影响）。在第三步中，对于在第二步中成功形成的每个组，使用单独的精子样本进行前瞻性 IVP。记录和比较每个组的卵裂、胚胎发育和囊胚率，并在各自特征组的高组和低组之间进行比较。令人惊讶的是，对单独影响的评估表明，MB、AI 和 MP 水平较低会导致胚胎发育和囊胚率较高（p<0.05），而卵裂率则没有观察到。我们得出结论，精子特征强烈影响体外受精（IVF）后的胚胎发育，影响受精卵达到囊胚阶段的能力。单独来看，MB、AI 或 MP 水平可能是定义当前胚胎生产系统中 IVP 效率的一些关键特征之一。
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