Human TIMP1 Is a Growth Factor That Improves Oocyte Developmental Competence.

Oocyte developmental competence is the ability of a mature oocyte to be fertilized and subsequently support embryonic development. Such competence is gained during folliculogenesis and is facilitated by the bidirectional communication into a compacted cumulus-oocyte complex (COC). Human tissue inhibitor of metalloproteinases-1 (TIMP1) participates in biological processes, including cell growth, differentiation, and apoptosis. This study aimed to evaluate the influence of TIMP1 as a growth factor on the in vitro maturation (IVM) culture of bovine COCs to improve oocyte developmental competence. All TIMP1 treatments (50, 100, and 150 ng/mL) favored the COCs' compaction structure (p < 0.05). TIMP1 at 150 ng/mL produced more oocytes in metaphase II compared to the other treatments (p < 0.05). The 150 ng/mL TIMP1 generated oocytes with the most (p < 0.05) cortical granules below the plasma membrane (pattern I). In a parthenogenesis assay, oocyte IVM in 50 ng/mL of TIMP1 produced the most blastocyst compared to the other treatments (p < 0.05). The Principal Component Analysis (PCA) showed that 50 ng/mL of TIMP1 was the best condition to develop oocyte competence because it was associated with the COC compact and cortical granule pattern I. TIMP1 influences the development of oocyte competence when added to the IVM culture medium of COCs.

IMD/ADM2 operates as a secretory factor that controls cumulus-oocyte complexes (COCs) conformation for oocytes in vitro maturation.

During in vitro maturation (IVM), the compact structure of cumulus-oocyte complexes (COCs) is vital for oocyte competence acquisition. Intermedin/Adrenomedullin-2 (IMD/ADM2) binds to the receptor RAMP (1, 2, or 3):CLR. Recently, it was demonstrated that IMD/ADM2 stimulates oocyte competence and improves bovine embryo quality. Therefore, this study aimed to examine the IMD/ADM2 as a secretory factor controlling COCs conformation for oocyte maturation. The results showed that traditional M-CDM medium induced in COCs the Imd/Adm2 gene expression during IVM and produced IMD/ADM2 peptide secretion. Furthermore, after IVM, in the oocytes, the expression of ramps (1, 2, or 3) and clr was demolished, and RAMPs and CLR proteins were decreased, with a negative Pearson correlation. These results suggest that RAMPs and CLR are synthesized and stored during oocyte maturation. Supplementing the M-CDM with α-RAMP1 or α-IMD/ADM2 antibodies elicits a negative effect (P < 0.05) in COCs compaction. Blocking the IMD/ADM2 signaling pathway with any α-RAMPs or α-CLR antibodies produces a similar lower yield of oocytes in metaphase II (P > 0.05) but was lower than control culture medium (P < 0.05). In conclusion, when COCs are cultured with M-CDM, the IMD/ADM2 becomes expressed and secreted. In turn, it acts as a ligand preferentially to RAMP1:CLR or RAMP3:CLR, present in cumulus cells and oocytes. Sequentially, COCs compact structure is conformed to promote an adequate bidirectional communication that conduces the oocytes' maturation.