A novel artificial oocyte activation method: using nanosecond pulsed electric fields (nsPEFs) to regulate calcium oscillation patterns

Abstract

KO and WT mice. 200 ml of each semen were used for total RNA extraction using the miRNeasy serum/plasma kit (Qiagen). RESULTS: Prmt5-and histone-associated protein Copr5 is highly expressed intestis and its depletion impacted on the maturation of spermatogonia to spermatids, although Copr5 KO animals were fertile. Extinction of Copr5 in mice testis leads to a down-regulation of the level of Miwi protein, as confirmed by western blot analyses and immunohistochemistry of WT and KO mice testis whole cell extracts and paraffin-embedded testis sections, respectively. In addition, the mRNA level of three pre-pachytene piR-NAs, prepiR1, prepiR2 and prepiR3 was decreased in KO Copr5 compared to WTmice, whereas in KO mice the expression level of the LINE1 mRNA, the most abundant retro-transposons that was usedas a read out of deregulation of the piRNA pathway, was increased. Using both human GEO data and patients with teratozoospermia, the present study reports a correlation between a low level of Copr5 mRNA and teratozoospermia, a human pathology characterized by abnormally shaped sperm that can negatively affect fertility. CONCLUSIONS: Copr5 KO in mice perturbed some genome surveillance mechanisms in germ cells by an alteration of the expression level of two major components present in the Piwi-interacting RNA (piRNA) pathway, the Miwi and the LINE1. In addition, low level of Copr5 mRNA correlates with human teratozoospermia sperm. Supported by: This work was supported by grants from the Ligue Contre le Cancer (CS) and the Association pour la Recherche contre le Cancer (EF).The authors declare no competing or financial interests.