Possible origin of glutamatergic projections to the midbrain periaqueductal gray and deep layer of the superior colliculus of the rat

The possible origin of glutamatergic input to the rodent periaqueductal gray (PAG) was analyzed utilizing a combined retrograde transport-immunocytochemical technique. Injections of wheat germ agglutinin-horseradish peroxidase were made into the PAG of 12 adult rats and into the deep layer of the superior colliculus in 2 rats. The brain tissue was first reacted histochemically to demonstrate the retrograde tracer and subsequently processed with immunohistochemical techniques using a recently developed monoclonal glutamate antibody. Following PAG injections, several brain areas were found to contain double-labeled neurons. The greatest number of double-labeled glutamate-like immunoreactive neurons were observed in the zona incerta, spinal trigeminal nucleus, cuneiform nucleus, cingulate cortex, cerebellar interpositus nucleus, deep mesencephalic nucleus and the PAG itself. Double-labeled neurons were also observed in several other nuclei including the pretectal nuclei, the frontal and occipital cortex, several reticular nuclei, the dorsomedial hypothalamic nucleus, and the substantia nigra. Many of the same nuclei contained double-labeled neurons following collicular injections, but in addition, double-stained cells were found in the primary visual cortex, lateral dorsal and lateral posterior thalamic nuclei, nucleus of the posterior commissure, ventral lateral geniculate nucleus, dorsal column nuclei and several additional pretectal nuclei. The results of mis double-labeling study raise the possibility that these nuclei may provide glutamatergic inputs to the midbrain PAG and/or superior colliculus. These putative glutamatergic afferent projections may ultimately influence the PAG's role in several important functions including antinociception, defensive mechanisms or vocalization and may also play a role in the superior collicular involvement in defensive mechanisms, in visuo-motor integration in the orienting reflex and in facilitating shifts in gaze.