“Relevance of the Cysteine Rich Secretory Protein (CRISP) family for female fertility

ABRIL REBAGLIATI CID,; VISACOVSKY, N; MARIANA WEIGEL MUÑOZ; VALERIA SULZYK; CUASNICÚ, PATRICIA S

Congreso; Reunión Anual de Sociedades de Biociencias; 2023

Cysteine-Rich Secretory Proteins (CRISP1-4) play key roles inmammalian fertilization in both sexes. In females, CRISP1, 2 and3 are expressed in reproductive organs and/or cumulus cells thatsurround the egg. Whereas single female knockout (KO) mice forCRISP1 and CRISP2 are fertile, double KO (DKO) females forCRISP1 and CRISP3 are subfertile, suggesting the existence ofcompensation among CRISP family members. To further investigatethe relevance of CRISP proteins for female fertility, triple KOfor CRISP1, 2 and 3 (TKO) and quadruple (QKO) were generatedby CRISPr/Cas9 and their fertility examined by caging one mutantand one control female with a fertile male, followed by evaluation ofborn pups. Results showed that fertility of TKO and QKO mice wassignificantly impaired compared to controls (p< 0.01) and not differentbetween mutant groups, consistent with the lack of CRISP4 infemales. As CRISP2/CRISP4 DKO females generated in our lab arefertile, fertility defects in TKO/QKO seem to be mainly due to the lackof CRISP1 and CRISP3. Based on this, in vivo fertilization studieswere performed in superovulated instead of estrous single CRISP1KO females to analyze the contribution of CRISP1 to female fertilityin a more demanding condition. Results showed a significantdecrease in the percentage of fertilized eggs recovered from theampulla, supporting the relevance of female CRISP1 for fertility. Asno single KO for CRISP3 was available at our lab, and with the aimof analyzing the contribution of CRISP3 to female fertility, femaleKO mice lacking this protein were generated by CRISPr-Cas9 andfertility analyzed. Differently from CRISP1 and CRISP2 single KOcolonies, a significant decrease in fertility was observed for CRISP3KO females. Together, these results support the relevance of femaleCRISP proteins, especially CRISP3, for fertility, contributing to a betterunderstanding of the molecular mechanisms underlying mammalianfemale fertility.