Rat small intestinal epithelium is equipped with peptide YY (PYY)-preferring receptors, which also recognize neuropeptide Y (NPY) with high affinity. We therefore examined the distribution of PYY-NPY receptors along the villus-crypt axis after separation of mature villus cells from proliferative crypt cells. Specific 125I-labeled PYY binding was nine times higher in crypt cells than in villus cells. This was not due to differential degradation of PYY or PYY binding sites by the two cell populations. Rather, Scatchard analysis of equilibrium binding data showed that binding capacity (Bmax) of receptors increased from villus to crypt. Bmax were 166 +/- 36 and 21 +/- 3 fmol/mg protein, and dissociation constants (Kd) were 0.10 +/- 0.02 and 0.05 +/- 0.02 nM in crude membranes prepared from crypt and villus cells, respectively. For all cell populations, NPY and rat pancreatic polypeptide were 8- and 1,800-fold less potent than PYY in inhibiting 125I-PYY binding, respectively. Therefore, receptors appear to be PYY preferring along the entire villus-crypt axis. Both peptides (at the maximally active concentration of 1 microM) reduced vasoactive intestinal peptide (VIP)-stimulated adenosine 3',5'-cyclic monophosphate (cAMP) by 50% in crypt cells. PYY was four to six times more potent than NPY in agreement with the expression of a PYY-preferring receptor. By contrast, neither PYY nor NPY altered VIP-stimulated cAMP levels in villus cells. These results indicate that PYY-preferring receptors, negatively coupled to the cAMP production system, are preferentially expressed in crypt cells where intestinal ionic secretion is believed to take place.