Our research delved into the exploration of spexin, a neuropeptide, within the stomach, small intestine, and colon of cows. While the roles of spexin in smaller animals are established, our focus was to bridge the knowledge gap regarding its functions in larger mammals. Through the analysis of selected segments of the unique gastrointestinal tract of the cattle, we sought to unravel the locations of spexin in the gastrointestinal tract of this polygastric animal. Remarkably, our findings revealed heightened gene expression of a specific spexin receptor, galanin receptor type 2, in the small intestine. This finding provides valuable insights into the functioning of spexin in larger livestock animals, particularly in terms of digestive regulation. The implications extend beyond veterinary considerations, potentially providing a window into understanding digestive processes in humans. This study enhances our overall understanding of neuropeptide functions and establishes a foundation for progress in animal health research.