The mammalian oviduct provides an optimal environment for the maturation of gametes, fertilization, and early embryonic development. Secretory cells lining the lumen of the mammalian oviduct synthesize and secrete proteins that have been shown to interact with and influence the activities of gametes and embryos. We hypothesized that the presence of gametes in the oviduct alters the oviductal secretory proteomic profile. We used a combination of two-dimensional gel electrophoresis and liquid chromatography-tandem mass spectrometry to identify oviductal protein secretions that were altered in response to the presence of gametes in the oviduct. The oviductal response to spermatozoa was different from its response to oocytes as verified by Western blotting. The presence of spermatozoa or oocytes in the oviduct altered the secretion of specific proteins. Most of these proteins are known to have an influence on gamete maturation, viability, and function, and there is evidence to suggest these proteins may prepare the oviductal environment for arrival of the zygote. Our findings suggest the presence of a gamete recognition system within the oviduct capable of distinguishing between spermatozoa and oocytes. The mammalian oviduct is the venue of important events leading to the establishment of pregnancy. These events include final maturation and transport of the female and male gametes, fertilization, cleavage-stage embryonic development, and transport of the embryo to the uterus. In mammals, the physiological interaction between gametes, embryos, and oviductal epithelia involves intimate and specific contact between the two cell types (1-6). During the estrous or menstrual cycle, the mammalian oviduct undergoes significant endocrine-induced morphological, biochemical, and physiological changes. These changes establish an essential microenvironment within the oviduct. Oviductal fluid is a crucial part of this milieu and consists of transudate from serum together with specific compounds synthesized by the luminal epithelium. The current dogma states that the oviductal environment and the composition of oviductal fluid are solely under the influence of the hormonal changes in the oviduct (7-9). However, in recent years, several investigations from our laboratory and others have challenged this view by providing evidence of transcriptional changes in the oviduct in response to gametes irrespective of oviductal hormonal status (10 -12).Although these data provide strong evidence in relation to the modulation of the oviductal environment by gametes, they lack information regarding changes to the oviductal proteomic profile, for example the secretory profile. In mammals, not all the changes in the transcriptome are translated into proteomic alterations due to post-translational modifications. Ellington et al. (13) and Thomas et al. (14) provide the only evidence that at least spermatozoa can influence the (secretory) proteomic profile of oviductal epithelial cells. These investigations have reported de novo protein synthesis...