Neural activity in the dorsal lateral geniculate nucleus of the thalamus (DLG) is modulated by an ascending cholinergic projection from the brainstem. The purpose of this study was to identify and localize specific muscarinic receptors for acetylcholine in the DLG. Receptors were identified in rat and cat tissue by means of antibodies to muscarinic receptor subtypes, ml-m4. Brain sections were processed immunohistochemically and examined with light and electron microscopy. Rat DLG stained positively with antibodies to the m1, m2,and m3 receptor subtypes but not with antibodies to the m4 receptor subtype. The m1 and m3 antibodies appeared to label somata and dendrites of thalamocortical cells. The m1 immunostaining was pale, whereas m3-positive neurons exhibited denser labeling with focal concentrations of staining. Strong immunoreactivity to the m2 antibody was widespread in dendrites and somata of cells resembling geniculate interneurons. Most m2-positive synaptic contacts were classified as F2-type terminals, which are the presynaptic dendrites of interneurons. The thalamic reticular nucleus also exhibited robust m2 immunostaining. Cat DLG exhibited immunoreactivity to the m2 and m3 antibodies. The entire DLG stained darkly for the m2 receptor subtype, except for patchy label in the medial interlaminar nucleus and the ventralmost C laminae. The staining for m3 was lighter and was distributed more homogeneously across the DLG. The perigeniculate nucleus also was immunoreactive to the m2 and m3 subtype-specific antibodies. Immunoreactivity in cat to the m1 or m4 receptor antibodies was undetectable. These data provide anatomical evidence for specific muscarinic-mediated actions of acetylcholine on DLG thalamocortical cells and thalamic interneurons.