PURPOSE. To determine the expression patterns of the vesicular acetylcholine transporter (VAChT), tyrosine hydroxylase (TH) and neuronal nitric oxide synthase (nNOS) in the pterygopalatine ganglion (PPG) and the exorbital lacrimal gland of normal mice. METHODS. Mouse PPG and lacrimal glands were processed for single- and double-labeled indirect immunofluorescence studies. Slides were examined with conventional fluorescence microscopy and confocal laser scanning microscopy. RESULTS. All the somata in the PPG expressed both VAChT and nNOS immunoreactivity (IR). The postganglionic axons within the ganglion showed less VAChT-immunoreactive intensity than that seen in the somata, whereas nNOS IR was almost undetectable. In the lacrimal gland, nNOS-positive nerve bundles and fibers were observed to be associated with tear-collecting ducts, blood vessels, and acini. Some nNOS-positive punctate elements appeared to be distributed among acini. Many nerve fibers were VAChT immunoreactive and a small number of fibers were TH immunoreactive in the gland. Most of the VAChT-positive fibers and some of the TH-positive nerves displayed nNOS IR. CONCLUSIONS. The expression of nNOS in cells of the PPG and in lacrimal gland nerves suggests that NO may play a role in modulating tear production. The site of action may include the PPG, ducts, blood vessels, acini, nerve fibers, and myoepithelial cells within the gland. NO may modulate parasympathetic and/or sympathetic synaptic transmission or by acting directly on lacrimal gland components. The interaction between NO-ergic and the conventional autonomic input illustrates the complexity of the innervation pattern of the mouse lacrimal gland.