Nutrient-evoked gastrointestinal reflexes are likely initiated by specialized epithelial cells located in the small intestine that detect luminal stimuli and release mediators that activate vagal endings. The G-protein, -gustducin, a key signal molecule in lingual taste detection, has been identified in mouse small intestine where it may also subserve nutrient detection; however the phenotype of -gustducin cells is unknown. Immunohistochemistry was performed throughout the mouse small intestine for -gustducin, enteroendocrine cell markers 5-HT and glucagon-like peptide-1 (GLP 1), and brush cell markers neuronal nitric oxide synthase (nNOS) and UEA-1 lectin binding, singly, and in combination. -gustducin was expressed in solitary epithelial cells of the mid-upper villus, which were distributed in a regional manner with most occurring within the mid-jejunum. Here, 27% of -gustducin cells co-labeled for 5-HT and 15% for GLP-1; 57% of -gustducin cells co-labeled UEA-1, with no triple labeling. -gustducin cells that co-labeled for 5-HT or GLP-1 were of distinct morphology and exhibited a different -gustducin immunolabeling pattern to those co-labeled with UEA-1. nNOS was absent from intestinal epithelium despite strong labeling in the myenteric plexus. We conclude that subsets of enteroendocrine cells in the mid-jejunum and brush cells (more generally distributed) are equipped to utilise -gustducin signaling in mice. Intestinal taste modalities may be signaled by these enteroendocrine cells via the release of 5 HT, GLP-1 or co-expressed mediators, or by brush cells via a non-nitrergic mediator in distinct regions of the intestine.