Durban Mercè, Barragán Montserrat, Colodron Marta, Ferrer-Buitrago Minerva, De Sutter Petra, Heindryckx Björn, Vernaeve Valérie, Vassena Rita
Clínica EUGIN, Travessera de les Corts 322, 08029, Barcelona, Spain.
J Assist Reprod Genet. 2015 Jun;32(6):879-86. doi: 10.1007/s10815-015-0496-0. Epub 2015 May 19.
Intracytoplasmic sperm injection (ICSI) is widely used to achieve fertilization in the presence of severe male factor, resulting in high fertilization rates. Nevertheless, 1-3 % of couples experience complete fertilization failure after ICSI. When a male factor is identified, assisted oocyte activation (AOA) can help overcome fertilization failures. The objective of this study is to describe a case of repeated complete fertilization failures after ICSI with donor oocytes, and to investigate the molecular and functional aspects of phospholipase C zeta (PLCζ) protein in the patient semen.
The patient was a normozoospermic male who had previously fathered, through natural conception, four children by a different partner. Molecular and functional analysis of sperm-specific PLCζ in the patient and control samples by means of gene sequencing, immunocytochemistry, Western blot, mouse oocyte activation test (MOAT), and mouse oocyte calcium analysis (MOCA) were used.
PLCζ expression levels and distribution were significantly disrupted, although MOAT and MOCA did not indicate a decrease in activation ability.
Normozoospermic males can have disrupted expression and distribution of PLCζ, and reduced activation ability after ICSI in human oocytes, despite their normal activation potential in functional testing using mouse oocytes. Discrepancy among molecular and functional data might exist, as mutations in the gene sequence may not be the only cause of alteration in PLCζ protein related to activation failures.
胞浆内单精子注射（ICSI）被广泛用于在严重男性因素存在的情况下实现受精，从而获得高受精率。然而，1%-3%的夫妇在ICSI后会经历完全受精失败。当确定存在男性因素时，辅助卵母细胞激活（AOA）有助于克服受精失败。本研究的目的是描述一例使用供体卵母细胞进行ICSI后反复出现完全受精失败的病例，并研究患者精液中磷脂酶Cζ（PLCζ）蛋白的分子和功能方面。
该患者为精子正常的男性，此前通过自然受孕与不同伴侣育有四个孩子。通过基因测序、免疫细胞化学、蛋白质印迹、小鼠卵母细胞激活试验（MOAT）和小鼠卵母细胞钙分析（MOCA）对患者和对照样本中的精子特异性PLCζ进行分子和功能分析。
尽管MOAT和MOCA未显示激活能力下降，但PLCζ的表达水平和分布明显紊乱。
精子正常的男性在使用人卵母细胞进行ICSI后，PLCζ的表达和分布可能会受到破坏，激活能力降低，尽管在使用小鼠卵母细胞的功能测试中其激活潜力正常。分子和功能数据之间可能存在差异，因为基因序列中的突变可能不是与激活失败相关的PLCζ蛋白改变的唯一原因。
