Tavalaee Marziyeh, Parivar Kazem, Shahverdi Abdol-Hossein, Ghaedi Kamran, Nasr-Esfahani Mohammad Hossein
a Department of Biology, School of Basic Science, Science and Research Branch , Islamic Azad University , Tehran , Iran.
b Department of Embryology, Reproductive Biomedicine Research Center , Royan Institute for Reproductive Biomedicine, ACECR , Tehran , Iran.
Hum Fertil (Camb). 2017 Jun;20(2):96-103. doi: 10.1080/14647273.2016.1264011. Epub 2017 Jan 12.
The aim of this study was to investigate the value of sperm factors PAWP and PLCζ involved in oocyte activation and sperm chromatin status, as indicators of sperm quality during uncapacitated, capacitated and acrosome-reacted conditions. Semen samples collected from 10 normozoospermic men and sperm factors PAWP, PLCζ, protamine deficiency and DNA fragmentation evaluated in fresh samples, processed sample by DGC (density gradient centrifugation), capacitated and acrosome reacted sperm, using immunofluorescence and flow cytometry. The status of capacitation and the acrosome reaction were confirmed by tyrosine kinase phosphorylation and CD46 markers, respectively. The proportion of sperm-expressing weak or zero levels of PLCζ and PAWP were significantly reduced after DGC processing. There were no significant differences among processed DGC, capacitated and acrosome-reacted samples. The proportions of DNA fragmented and protamine-deficient spermatozoa were decreased after DGC and maintained during capacitation and the acrosome reaction. DGC processing, therefore, improved the quality of sperm in terms of chromatin and DNA integrity, and removed sperm which may have had a lower potential to induce oocyte activation and undergo capacitation and the acrosome reaction.
本研究的目的是探讨精子因子PAWP和PLCζ在卵母细胞激活及精子染色质状态方面的价值，以此作为未获能、获能及顶体反应状态下精子质量的指标。从10名正常精子男性收集精液样本，并使用免疫荧光和流式细胞术对新鲜样本、经密度梯度离心（DGC）处理的样本、获能精子及顶体反应精子中的精子因子PAWP、PLCζ、鱼精蛋白缺乏情况和DNA片段化进行评估。分别通过酪氨酸激酶磷酸化和CD46标记物确认获能状态和顶体反应状态。经DGC处理后，表达PLCζ和PAWP水平较弱或为零的精子比例显著降低。经DGC处理的样本、获能样本和顶体反应样本之间无显著差异。经DGC处理后，DNA片段化精子和鱼精蛋白缺乏精子的比例降低，并在获能和顶体反应过程中保持。因此，DGC处理在染色质和DNA完整性方面改善了精子质量，并去除了可能诱导卵母细胞激活、获能及顶体反应能力较低的精子。
