IBYME   02675
INSTITUTO DE BIOLOGIA Y MEDICINA EXPERIMENTAL
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Characterization of CRISP4 Knockout mice.
Autor/es:
CARVAJAL G; WEIGEL MUÑÓZ M; BRUKMAN NG; IKAWA M; OKABE M; CUASNICÚ PS
Lugar:
Chascomús
Reunión:
Jornada; XVI Jornadas de la Sociedad Argentina de Biología; 2014
Institución organizadora:
Sociedad Argentina de Biología (SAB)
Resumen:
Epididymal CRISP1 and CRISP4 proteins are present in sperm and reported to be involved in gamete fusion and sperm zona pellucida (ZP) binding, respectively. However, knockout (KO) mice for these molecules are fertile, suggesting compensatory mechanisms between homologous proteins. With the aim of developing a double KO mice, and based on the availability of CRISP1 KO mice in our laboratory, we generated and characterized a colony of CRISP4 KO mice. RT-PCR and Western blot confirmed the lack of Crisp4 mRNA in the epididymis and of CRISP4 protein in both epididymis and sperm, as well as normal expression of CRISP1. Whereas no differences in fertility, in vivo fertilization, sperm number, motility or viability between wildtype (WT), heterozygote (HT) and KO mice were observed, HT and KO spermatozoa exhibited lower levels of progesterone-induced acrosome reaction (AR) than WT cells. Consistent with this, a lower expression of CRISP4 was found in the epididymis of HT vs WT. Exposure of fresh and capacitated sperm to different treatments showed that CRISP4 was not released by NaCl 0,6M or 5U/mL PLC-PI but was completely removed by Tritón X-100, indicating a strong association of CRISP4 with capacitated sperm. This, together with the correlation between CRISP4 presence in sperm and their ability to undergo the AR, supports a role for CRISP4 in additional stages of fertilization such as cumulus and ZP penetration. This, and CRISP1/CRISP4 KO mice fertility are at present under investigation.