Claudins (CLDNs) are major Ca2+-independent cell adhesion molecules functioning at tight junctions (TJ). The presence and localization of cell adhesion molecules are important for understanding the mechanisms associated with follicular and luteal development in the ovary. In this study, there was an examination of whether CLDN-1 and CLDN-5 are present in a cell- and stage-specific manner during follicular and luteal development in the domestic cat ovary using immunohistochemistry and Western blot analysis. While results from immunoblot analyses revealed there were relatively similar abundances of CLDN-5 protein in three phases of the ovarian cycle, the abundance of CLDN-1 in the luteal phase was greater than those measured in the follicular and anestrous phases (P < 0.01). Results with immunohistochemistry indicate CLDN-1 and -5 are mainly localized in the cell nuclei and cytoplasm of all tissues of the cat ovary. In follicles, throughout the development from primordial to large antral follicles, CLDN-1 and -5 were present in oocytes, and the granulosa and theca cell layers. In follicles at all stages of atresia, there were cell-type and stage-specific protein distributions with immunostaining present in granulosa, thecal interstitial, and fibroblast-like cells. In corpora lutea, both small and large luteal cells stained positively for both claudins. In conclusion, the specific presence and localization patterns of CLDN-1 and -5 in the cat ovary is suggestive that these TJ proteins could have local functions in the regulation of most ovarian functions such as follicle development and atresia, ovulation, and corpus luteum formation and regression.