A new role for crisp1 in the fertilization process

ERNESTO JI; BATTISTONE MA; CALB D; WEIGEL MUÑOZ M; COHEN DJ ; CUASNICU PS

New Hampshire

Conferencia; Gordon Research Conference; 2011

Epididymal protein CRISP1 is the first described member of family reported to be present mainly in the male reproductive tract. Evidence from our group, obtained using in vitro fertilization assays and knock out mice generated in our laboratory, support the participation of CRISP1 in sperm capacitation as well as in sperm interaction with both the zona pellucida (ZP) and the oolema.  In the present work, the CRISP1 knock out (KO) mice were used to investigate the presence of the protein in the female reproductive tract and its potential role in the fertilization process. Rt-PCR analysis revealed the presence of CRISP1 mRNA in the uterus, ovary, oviduct, and cumulus cells of wild type (WT) but not KO mice. Western blot (Wb) and indiirect immunofluorescence studies using a specific polyclonal anti-CRISP1 antibody confirmed the presence of the protein only in the tissues and cumulus cells from WT females. In view of these observations, we evaluated the possible participation of female CRISP1 in fertilization by in vitro fertilization assays performed using cumuls-oocyte complexes (COCs) and sperm from WT and KO mice. Results showed that when WT sperm were used for insemination, significantly lower levels of fertilization were observed for KO than for WT COC. This decrease was even more evident when KO COCs were exposed to KO instead than to WT sperm. Together, these results confirm the existence of CRISP1 in the female reproductive tract and support the existence of a new role for CRISP1 in the fertilization process