Vasoactive intestinal peptide (VIP) is an endogenous neuropeptide with a broad array of physiological functions in many organs including the intestine. Its actions are mediated via G- protein coupled receptors and VPAC1 is the key receptor responsible for majority of VIP's biological activity. The distribution of VPAC1 along the length of the gastrointestinal tract and its sub cellular localization in intestinal epithelial cells has not been fully characterized. The current studies were undertaken to determine VPAC1 distribution and localization so that VIP based therapies can be targeted to specific regions of the intestine. The results indicated that the mRNA levels of VPAC1 showed an abundance pattern of colon> ileum> jejunum in the mouse intestine. In parallel, the VPAC1 protein levels were higher in the mouse colon, followed by the ileum and jejunum. Immuno-fluorescence studies in mouse colon demonstrated that the receptor was specifically localized to the luminal surface as evident by co-localization with the apical marker villin but not with the basolateral marker Na⁺/K⁺ ATPase. In the human intestine, VPAC1 mRNA expression exhibited a distribution similar to mouse intestine and was highest in the sigmoid colon. Furthermore, in the human colon, VPAC1 also showed predominantly apical localization. The physiological relevance of the expression and apical localization of VPAC1 remains elusive. We speculate that apical VPAC1 in intestinal epithelial cells may have relevance in recognizing secreted peptides in the intestinal lumen and therefore, supports the feasibility of potential therapeutic and targeting use of VIP formulations via oral route to treat GI diseases.