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During the estrous cycle, uterine expression of NCKX3 mRNA and protein was enhanced up to 4.0- and 2.5-fold, respectively, at proestrus compared to during estrus and diestrus. To examine the effect of sex steroids on NCKX3 regulation in the uterus, immature female rats were treated with E2 (40??g/kg body weight; BW), P4 (4?mg/kg BW), or E2 plus P4 for 3 days. The expression of NCKX3 Temozolomide manufacturer mRNA and protein was induced by E2, whereas P4 antagonized E2-induced NCKX3 expression. Subsequent immunohistochemical analysis revealed that uterine NCKX3 protein was abundantly localized in the cytoplasm of luminal and glandular epithelial cells throughout the estrous cycle. Taken together, these results indicate that uterine NCKX3 is abundantly expressed in the uterus and that its expression is regulated by the steroid hormones, E2 and P4. These findings suggest that NCKX3 may be involved in reproductive function during the estrous cycle in female rats. Mol. Reprod. Dev. 77:971�C977, 2010. ? 2010 Wiley-Liss, Inc. ""Recent studies have independently shown that cyclic DDR1 adenosine 3��5��-monophosphate (cAMP) modulation prior to in vitro maturation (IVM) and epidermal growth factor (EGF)-like peptide supplementation during IVM improve subsequent oocyte developmental outcomes. This study investigated the effects of an IVM system that incorporates these two concepts. Cumulus�Coocyte complexes (COCs) were collected from pre-pubertal mice either 46?hr post-equine chorionic gonadotropin (eCG) (IVM) or post-eCG?+?post-human chorionic gonadotropin (hCG) stimulation (in vivo maturation; IVV). IVM COCs were treated with the cAMP modulators forskolin and IBMX for 1, 2, or 4?hr (pre-IVM phase) prior to IVM. COCs then underwent IVM with the EGF-like peptides amphiregulin or epiregulin, or with the common IVM stimulants follicle-stimulating hormone (FSH) or EGF. A pre-IVM phase increased the size of the subsequent blastocysts' inner-cell-mass compared to standard IVM, regardless of IVM treatment (P?selleck kinase inhibitor significantly increased blastocyst quality (trophectoderm and total cell numbers) and/or yield (P?