We have previously demonstrated that a variant form of choline acetyltransferase (pChAT) is expressed in rat trigeminal neurons. To assess the significance of pChAT in sensory functions, we characterized immunohistochemically pChAT-positive trigeminal neurons in the rat. pChAT-immunoreactivity was observed in a rather uniform pattern in about half of all trigeminal neurons throughout the trigeminal ganglion. The majority of pChAT-positive neurons had small to medium-sized cell bodies. Double immunofluorescent study showed that more than 90% of substance P (SP)-positive trigeminal cells and about 80% of calcitonin gene-related peptide (CGRP)-positive cells exhibited pChAT-immunoreactivity. pChAT-positive cells formed a larger population of neurons than SP-positive or CGRP-positive cells, but they were a different population from calbindin-D(28k)-positive neurons. In addition, pChAT-immunoreactivity was present in a subset of neurons positive for neuronal nitric oxide synthase. The present results suggest that pChAT plays roles not only in nociception, but also in other sensory functions such as mechanoreception mediating tactile sensation.