Identification of rat brainstem multisynaptic connections to the oral motor nuclei using pseudorabies virus III. Lingual muscle motor systems

The present experiments complete our investigations of higher order afferent control of the orofacial muscles by examining the premotor systems controlling the lingual musculature. Pseudorabies virus (PRV) was injected into the extrinsic (protruders: genioglossus and geniohyoid; retractors: hyoglossus and styloglossus) and intrinsic tongue muscles in bilaterally sympathectomized rats. Injection volumes ranged from 1 to 12 μl with average titers of 4 X 10⁸ pfu/ml and maximum survival times of 90 h. Consistent labeling patterns and distributions occurred across each of the individual muscles and between extrinsic and intrinsic muscle groups, as well as in comparison to the results from the previous masticatory and facial muscle experiments. Virus injections produced a predictable myotopic labeling pattern in the hypoglossal nucleus (Mo 12). Transneuronally labeled neurons occurred in regions known to project directly to Mo 12 moroneurons including the nucleus subcoeruleus, trigeminal sensory areas, parvicellular reticular formation, and the dorsal medullary reticular fields. Maximum survival times revealed more distant connections from medial and lateral reticular zones including the periaqueductal gray, dorsal raphe, laterodorsal and pedunculopontine tegmental areas, and substantia nigra in the midbrain, the gigantocellular region, pontine nucleus caudalis and ventralis, and lateral paragigantocellular region in the pons, and the nucleus of the solitary tract, paratrigeminal region, and paramedian field in the medulla. Thus, injections of PRV into the orofacial muscles revealed a complex, but remarkably uniform network of multisynaptic connections in the brainstem that control and coordinate the activity of the masticatory, facial, and lingual muscles.