Department of Reproductive Medicine, Ghent University Hospital, De Pintelaan 185, B-9000 Ghent, Belgium.
Reprod Biomed Online. 2010 Dec;21(6):742-9. doi: 10.1016/j.rbmo.2010.07.006. Epub 2010 Jul 18.
Failed fertilization after intracytoplasmic sperm injection (ICSI) can be due to a reduced oocyte-activation capacity caused by reduced concentrations and abnormal localization of the oocyte-activation factor phospholipase C (PLC) zeta. Patients with this condition can be helped to conceive by artificial activation of oocytes after ICSI with calcium ionophore (assisted oocyte activation; AOA). However some concern still exists about this approach. Mouse models could help to identify potential oocyte-activation strategies and evaluate their safety. In this study, the fertilizing capacity of wobbler sperm cells was tested and the efficiency of AOA with two exposures to ionomycin to restore fertilization and embryo development was studied. The quality of the obtained blastocysts was assessed and embryo transfer was performed to evaluate post-implantation development. The presence of PLCzeta in the spermatozoa and testis of the wobbler mouse was evaluated by PLCzeta immunostaining and quantitative reverse-transcription polymerase chain reaction. Sperm cells from wobbler mice had reduced fertilizing capacity and abnormalities in PLCzeta localization, but not in its expression. Artificially activating the oocytes restored fertilization and embryo development. Therefore, the wobbler mouse can be a model for failed fertilization after ICSI to study PLCzeta dynamics and aid in optimization of the AOA method.
卵胞浆内单精子注射(ICSI)后受精失败可能是由于卵激活因子磷脂酶 C(PLC)ζ的浓度降低和异常定位导致卵激活能力降低所致。通过在 ICSI 后用钙离子载体(辅助卵激活;AOA)人工激活卵,可以帮助患有这种疾病的患者受孕。然而,人们对此方法仍存在一些担忧。小鼠模型可以帮助确定潜在的卵激活策略并评估其安全性。在这项研究中,测试了 wobbler 精子细胞的受精能力,并研究了用两次离子霉素暴露来恢复受精和胚胎发育的 AOA 的效率。评估了获得的囊胚的质量,并进行胚胎移植以评估植入后的发育情况。通过 PLCzeta 免疫染色和定量逆转录聚合酶链反应评估 wobbler 小鼠精子和睾丸中 PLCzeta 的存在。 wobbler 小鼠的精子细胞受精能力降低,PLCzeta 定位异常,但表达正常。人工激活卵恢复了受精和胚胎发育。因此, wobbler 小鼠可以作为 ICSI 后受精失败的模型,用于研究 PLCzeta 动力学并有助于优化 AOA 方法。