The aim of this study was to map the viscerotopic representation of the upper alimentary tract in the sensory ganglia of the IXth and Xth cranial nerves and in the subnuclei of the solitary and spinal trigeminal tracts. Therefore, in 172 rats 0.5-65 microliters of horseradish peroxidase (HRP), wheat germ agglutinin-HRP, or cholera toxin-HRP were injected into the trunks and major branches of the IXth and Xth cranial nerves as well as into the musculature and mucosa of different levels of the upper alimentary and respiratory tracts. The results demonstrate that the sensory ganglia of the IXth and Xth nerves form a fused ganglionic mass with continuous bridges of cells connecting the proximal and distal portions of the ganglionic complex. Ganglionic perikarya were labeled in crude, overlapping topographical patterns after injections of tracers into nerves and different parts of the upper alimentary tract. After injections into the soft palate, pharynx, esophagus, and stomach, anterograde labeling was differentially distributed in distinct subnuclei in the nucleus of the tractus solitarius (NTS). Palatal and pharyngeal injections resulted primarily in labeling of the interstitial and intermediate subnuclei of the NTS and in the paratrigeminal islands (PTI) and spinal trigeminal complex. Esophageal and stomach wall injections resulted in labeling primarily of the subnucleus centralis and subnucleus gelatinosus, respectively. The distribution of upper alimentary tract vagal-glossopharyngeal afferents in the medulla oblongata has two primary groups of components, i.e., a viscerotopic distribution in the NTS involved in ingestive and respiratory reflexes and a distribution coextensive with fluoride-resistant acid-phosphatase-positive regions of the PTI and spinal trigeminal nucleus presumably involved in visceral reflexes mediated by nociceptive or chemosensitive C fibers.

We employed the neural tracers cholera toxin-horseradish peroxidase and wheat germ agglutinin-horseradish peroxidase to examine the organization of the afferent and efferent connections of the stomach within the medulla oblongata of the rat. The major finding of this study is that gastric motoneurons of the dorsal motor nucleus (DMN) possess numerous dendrites penetrating discrete regions of the overlying nucleus of the solitary tract (NTS). In particular, dendritic labelling was present in areas of NTS which also received terminals of gastric vagal afferent fibers such as the subnucleus gelatinosus, nucleus commissuralis, and medial nucleus of NTS. This codistribution of afferent and efferent elements of the gastric vagus may provide loci for monosynaptic vagovagal interactions. A small number of dendrites of DMN neurons penetrated the ependyma of the fourth ventricle and a few others entered the ventral aspect of the area postrema, thus making possible the direct contact of preganglionic neurons with humoral input from the cerebrospinal fluid and/or the peripheral plasma. Nucleus ambiguus neurons projecting to the stomach predominantly innervate the forestomach. The dendrites of these cells, when labelled, were generally short, and extended beyond the compact cluster of ambiguus neurons in a ventrolateral direction, parallel to the fascicles of vagal efferent fibers traversing the medulla.

Uptake, replication, and transneuronal passage of a swine neurotropic herpesvirus (pseudorabies virus, PRV) was evaluated in the rat CNS. PRV was localized in neural circuits innervating the tongue, stomach, esophagus and eye with light microscopic immunohistochemistry. In each instance, the distribution of PRV-immunoreactive neurons was entirely consistent with that observed following injection of cholera toxin-horseradish peroxidase conjugate (CT-HRP). Injections of the tongue resulted in retrograde transport of PRV and CT-HRP to hypoglossal motor neurons, while preganglionic neurons in the dorsal motor vagal nucleus or somatic motor neurons in the nucleus ambiguus were labeled following injections of the stomach or esophagus, respectively. At longer times after infection, viral antigens were found in astrocytes adjacent to infected neurons and their efferent axons and second-order neuron labeling became apparent. The distribution of second-order neurons was also entirely dependent upon the site of PRV injection. Following tongue injection, second-order neurons were observed in the trigeminal complex, the brain-stem tegmentum and in monoaminergic cell groups. Injection of the stomach or esophagus led to second-order neuron labeling confined to distinct subdivisions of the neucleus of the solitary tract and monoaminergic cell groups. Comparative quantitative analysis of the number of PRV immunoreactive neurons present in the diencephalon and brain stem following injection of virus into both the eye and stomach musculature of the same animal demonstrated that retrograde transport of PRV from the viscera was more efficient and occurred at a much faster rate than anterograde transport of virus. These data demonstrate projection-specific transport of PRV in the nervous system and provide further insight into the means through which this neurotropic virus infects the nervous system.

Pseudorabies virus (PRV) has been used extensively to map synaptic circuits in the CNS and PNS. A fundamental assumption of these studies is that the virus replicates within synaptically linked populations of neurons and does not spread through the extracellular space or by cell-to-cell fusion. In the present analysis we have used electron microscopy to characterize pathways of viral replication and egress that lead to transneuronal infection of neurons, and to document the non-neuronal response to neuronal infection. Three strains of PRV that differ in virulence were used to infect preganglionic motor neurons in the dorsal motor nucleus of the vagus (DMV). The data demonstrate that viral replication and transneuronal passage occur in a stepwise fashion that utilizes existing cellular processes, and that the non-neuronal response to infection serves to restrict nonspecific spread of virus by isolating severely infected neurons. Specifically, capsids containing viral DNA replicate in the cell nucleus, traverse the endoplasmic reticulum to gain access to the cytoplasm, and acquire a bilaminar membrane envelope from the trans cisternae of the Golgi. The outer leaf of this envelope fuses with the neuron membrane to release virus adjacent to axon terminals that synapse upon the infected cell. A second fusion event involving the viral envelope and the afferent terminal releases the naked capsid into the bouton. Systematic analysis of serial sections demonstrated that release of virus from infected neurons occurs preferentially at sites of afferent contact. Nonspecific diffusion of virus from even the most severely infected cells is restricted by astrocytes and other non-neuronal elements that are mobilized to the site of viral infectivity. The ability of glia and macrophages to restrict spread of virus from necrotic neurons is the product of (1) temporal differences in the mobilization of these cells to the site of infection, (2) differential susceptibility of these cells to PRV infection, and (3) abortive viral replication in cells that are permissive for infection. The findings provide further insight into the intracellular routes of viral assembly and egress and support the contention that transneuronal spread of virus in the brain results from specific passage of virions through synaptically linked neurons rather than through cell fusion or release of virus into the extracellular space.

We applied the neuroanatomical tracers cholera toxin-horseradish peroxidase and wheat germ agglutinin-horseradish peroxidase to investigate the neural connections of the area postrema (AP) in the rat. We find that the AP projects to the nucleus of the solitary tract (NTS) and dorsal motor nucleus of the vagus bilaterally both rostral and caudal to obex; the nucleus ambiguus; the dorsal aspect of the spinal trigeminal tract and nucelus and the paratrigeminal nucleus; the region of the ventrolateral medullary catecholaminergic column; the cerebellar vermis; and a cluster of structures in the dorsolateral pons which prominently include a discrete set of subnuclei in the lateral parabrachial nucleus. The major central afferent input to the area postrema is provided by a group of neurons in the paraventricular and dorsomedial hypothalamic nuclei whose collective dendrites describe a horizontally oriented plexus which encircles the parvocellular nucleus of the hypothalamus bilaterally. In addition, the caudal NTS may project lightly to the AP. The lateral parabrachial nucleus provides a very light input as well. These connections, when considered in the context of the known vagal afferent input and reduced blood-brain barrier of AP, place this structure in a unique position to receive and modulate ascending interoceptive information and to influence autonomic outflow as well.