ABSTRACT: This study aims to uncover how extracellular vesicles (EVs) influence key reproductive processes by isolating, characterizing, and analyzing the miRNA cargo of these oviductal and uterine fluid-EVs. Small RNA sequencing identified 388 miRNAs in OF-EVs and UF-EVs. However, 4 miRNAs significantly more abundant in OF-EVs, while 17 were significantly more abundant in UF-EVs, indicating potential differences in their roles in embryo-maternal communication. The most abundant miRNAs in both fluids included ocu-miR-148a-3p, ocu-let-7i-5p, and ocu-miR-10b-5p, which are associated with embryo development and implantation. Gene target prediction analysis of the top abundant revealed that miRNAs in OF-EVs regulate key signaling pathways such as MAPK, Hippo, TGF-β, Ras, PI3K-Akt, and mTOR, while UF-EVs are associated with Hippo, PI3K-Akt, FoxO, endocytosis, and mTOR signaling. These pathways are essential for cell proliferation, differentiation, and immune regulation, crucial for embryo-maternal interactions. Our findings suggest that EVs in rabbit reproductive fluids play a fundamental role in early embryonic development and successful pregnancy. These results provide new insights into reproductive biotechnology and veterinary medicine, with potential applications in improving fertility and embryo viability.