The purpose of the present study was to obtain reliable evidence for the presence of gamma-aminobutyric acid-ergic (GABAergic) synapses upon mesencephalic trigeminal nucleus (MTN) neurons in the rat. For confocal laser-scanning microscopy, phosphate-activated glutaminase-like immunoreactivity (-IR) of MTN neurons was visualized with red fluorescence of Texas Red, while glutamic acid decarboxylase (GAD)-IR of GABA axons was visualized with green fluorescence of dichlorotriazinyl aminofluorescein. Many GAD-axon terminals were in close apposition to the cell bodies of MTN neurons. For electron microscopy, MTN neurons were labeled with wheat germ agglutinin-horseradish peroxidase injected into the masseter nerve, while axon terminals were labeled with GAD-IR. GAD-axon terminals were in symmetric synaptic contact with the cell bodies of MTN neurons. Primary proprioceptive neurons in the orofacial regions might be regulated post-synaptically by GABA neurons.