Increasingly strong evidence suggests that cholinergic neurons in the mesopontine tegmentum play important roles in the control of wakefulness and sleep. To understand better how the activity of these neurons is regulated, the potential afferent connections of the laterodorsal (LDT) and pedunculopontine tegmental nuclei (PPT) were investigated in the rat. This was accomplished by using retrograde and anterograde axonal transport methods and NADPH-diaphorase histochemistry. Immunohistochemistry was also used to identify the transmitter content of some of the retrogradely identified afferents. Following injections of the retrograde tracer wheatgerm agglutinin-conjugated horseradish peroxidase (WGA-HRP) into either the LDT or the PPT, labelled neurons were seen in a number of limbic forebrain structures. The medial prefrontal cortex and lateral habenula contained more retrogradely labelled neurons from the LDT, whereas in the bed nucleus of the stria terminalis and central nucleus of the amygdala, more cells were labelled from the PPT. Moderate numbers of neurons were seen in the magnocellular regions of the basal forebrain, and many labelled neurons were observed in the lateral hypothalamus, the zona incerta, and the midbrain central gray from both the LDT and the PPT. Accessory oculomotor nuclei in the midbrain as well as eye movement-related structures in the lower brainstem contained some neurons labelled from the LDT, and fewer neurons from the PPT. A few labelled neurons were seen in somatosensory and other sensory relay nuclei in the brainstem and the spinal cord. Retrograde labelling was seen in a number of extrapyramidal structures, including the globus pallidus, entopenduncular and subthalamic nuclei, and substantia nigra following PPT injections; with LDT injections, labelling was similar in density in the substantia nigra but virtually absent in the entopeduncular and subthalamic nuclei. Data with the fluorescent retrograde tracer fluorogold combined with immunofluorescence indicated that many neurons in the zona incerta-lateral hypothalamic region that were retrogradely labelled from the LDT contained alpha-melanocyte-stimulating hormone. Numerous neurons were labelled throughout the reticular formation of the brainstem following either LDT or PPT injections. Many neurons retrogradely labelled in the LDT and PPT, the dorsal and median raphe nuclei, and the locus ceruleus contained choline acetyltransferase, serotonin, and tyrosine hydroxylase, respectively. The anterograde tracers WGA-HRP and phaseolus vulgaris leucoagglutinin were used to confirm some of the projections indicated by the retrograde labelling data; anterograde labelling was seen in the LDT and PPT following injections of one of these tracers into the medial prefrontal cortex, lateral hypothalamus, and the contralateral LDT.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent connections of the laterodorsal and the pedunculopontine tegmental nuclei in the rat: A retro‐ and antero‐grade transport and immunohistochemical study

The pedunculopontine tegmental nucleus (PPTn) was originally defined on cytoarchitectonic grounds in humans. We have employed cytoarchitectonic, cytochemical, and connectional criteria to define a homologous cell group in the rat. A detailed cytoarchitectonic delineation of the mesopontine tegmentum, including the PPTn, was performed employing tissue stained for Nissl substance. Choline acetyltransferase (ChAT) immunostained tissue was then analyzed in order to investigate the relationship of cholinergic perikarya, dendritic arborizations, and axonal trajectories within this cytoarchitectonic scheme. To confirm some of our cytoarchitectonic delineations, the relationships between neuronal elements staining for ChAT and tyrosine hydroxylase were investigated on tissue stained immunohistochemically for the simultaneous demonstration of these two enzymes.



The PPTn consists of large, multipolar neurons, all of which stain immunohistochemically for ChAT. It is present within cross-sections that also include the A-6 through A-9 catecholamine cell groups and is traversed by catecholaminergic axons within the dorsal tegmental bundle and central tegmental tract. The dendrites of PPTn neurons respect several nuclear boundaries and are oriented perpendicularly to several well-defined fiber tracts. Cholinergic axons ascend from the mesopontine tegmentum through the dorsal tegmental bundle and a more lateral dorsal ascending pathway. A portion of the latter terminates within the lateral geniculate nucleus.



It has been widely believed that the PPTn is reciprocally connected with several extrapyramidal structures, including the globus pallidus and substantia nigra pars reticulata. Therefore, the relationships of pallidotegmental and nigrotegmental pathways to the PPTn were investigated employing the anterograde autoradiographic methodology. The reciprocity of tegmental connections with the substantia nigra and entopeduncular nucleus was investigated employing combined WGA-HRP injections and ChAT immunohistochemistry.



The pallido- and nigrotegmental terminal fields did not coincide with the PPTn, but, rather, were located just medial and dorsomedial to it (the midbrain extrapyramidal area). The midbrain extrapyramidal area, but not the PPTn, was reciprocally connected with the substantia nigra and entope-duncular nucleus. We discuss these results in light of other cytoarchitec-tonic, cytochemical, connectional, and physiologic studies of the functional anatomy of the mesopontine tegmentum.

Pedunculopontine tegmental nucleus of the rat: cytoarchitecture, cytochemistry, and some extrapyramidal connections of the mesopontine tegmentum.

Projections from the auditory thalamus to the amygdala have been implicated in the processing of the emotional signficance of auditory stimuli. In order to further our understanding of the contribution of thalamoamygdala projections to auditory emotional processing, acoustic response properties of single neurons were examined in the auditory thalamus of chloral hydrate-anesthetized rats. The emphasis was on the medial division of the medial geniculate body (MGm), the suprageniculate nucleus (SG), and the posterior intralaminar nucleus (PIN), thalamic areas that receive inputs from the inferior colliculus and project to the lateral nucleus of the amygdala (AL). For comparison, recordings were also made from the specific thalamocortical relay nucleus, the ventral division of the medial geniculate body (MGv). Responses latencies were not statistically different in MGv, MGm, PIN, and SG, but were longer in the posterior thalamic region (PO). Overall, frequency tuning functions were narrower in MGv than in the other areas but many cells in MGm were as narrowly tuned as cells in MGv. There was some organization of MGv, with low frequencies represented dorsally and high frequencies ventrally. A similar but considerably weaker organization was observed in MGm. While the full range of frequencies tested (1–30 kHz) was represented in MGv, cells in MGm, PIN, and SG tended to respond best to higher frequencies (16–30 kHz). Thresholds were higher in PIN than in MGv (other areas did not differ from MGv). Nevertheless, across the various areas, the breadth of tuning was inversely related to threshold, such that more narrowly tuned cells tended to have lower thresholds. Many of the response properties observed in MGm, PIN, and SG correspond with properties found in AL neurons and thus add support to the notion that auditory responses in AL reflect thalamoamygdala transmission.

Response properties of single units in areas of rat auditory thalamus that project to the amygdala. II. Cells receiving convergent auditory and somatosensory inputs and cells antidromically activated by amygdala stimulation.

Projections to the amygdala from the auditory thalamus have been implicated in the associative conditioning of fear responses to acoustic stimuli. Thalamo-amygdala auditory projections enter the amygdala via the lateral nucleus (LA). It is well documented that these projections originate in the medial division (MGm) of the medial geniculate nucleus (MGN), the posterior intralaminar nucleus (PIN), and the suprageniculate nucleus (Sg). It is not known, however, whether these thalamic projections terminate in a topographic fashion within the LA. We therefore used several retrograde tract tracing techniques to determine whether the terminations of thalamo-amygdala fibers have a topographic organization within the LA. These tracers were injected into various locations within the LA, and the distribution of the retrogradely labeled cells throughout the thalamus was analyzed. In general, rostral to caudal distinctions in the thalamus are maintained in the LA, such that projections from throughout the MGN terminate in the anterior part of the LA, whereas the caudal part of the MGN projects to the caudal part of the LA. Furthermore, the density of cells that give rise to thalamo-amygdala projections varies within each thalamic nucleus along the rostro-caudal axis. The patterns of thalamo-amygdala connectivity observed support previous parcellation schemes that segregate the LA into dorsal, medial, and lateral areas, and suggest that the LA should be further divided into anterior and posterior parts. In addition to the well-known projections to the LA originating from PIN, MGN, and Sg, we also found substantial projections from the dorsal portion of the MGN (MGd) and the lateral posterior thalamic nucleus (LP). These findings suggest that some of the functional segregation in the thalamus may be preserved in the LA, and that the role of the MGd and LP in thalamo-amygdala transmission should be reconsidered.

Organization of projections to the lateral amygdala from auditory and visual areas of the thalamus in the rat

Tactile responses in the granule cell layers of the cerebellar hemispheres of rats are topographically arranged as a series of patches each representing a different region of the body surface. Previous observations had suggested that patches representing specific body parts recur in similar folial positions in different individuals; however, these relationships were not quantified. In this study we make inter-animal comparisons of the detailed distribution of receptive fields in the granule cell layer of the crown of crus IIa by using physiological mapping techniques. The results suggest that maps from different individuals do, in fact, share several topological features. These include the regions of the body surface represented, the general proportions of these representations, the relative positions of patches representing the same body parts, and the organization of receptive fields within patches located in similar positions. The principal variability seen in these comparisons was in the detailed neighborhood relations between different patches. As a result of the analysis of the consistent and variable features of these maps, we propose and discuss a new role for these cerebellar regions in coordinating the acquisition of tactile sensory information.

Variability in tactile projection patterns to cerebellar folia crus IIA of the Norway rat.

Projection systems from the gracile nucleus and the cuneate nuclear complex to their terminal sites in the mesencephalon, diencephalon, and cerebellum were examined by means of anterograde autoradiography and retrograde horseradish peroxidase methods.



Three projection systems emerge from the dorsal column nuclei, decussate via internal arcuate fibers, and form the contralateral medial lemniscus (ML). At the obex, some fibers split off the ML and course dorsolaterally, forming an ascending lateral system which fits the “lemniscal adjunct channel” (LAC) concept of Graybiel ('72). The ML continues rostrally as the “main lemniscal line channel” (MLLC). At the inferior colliculus, some LAC fibers terminate in the pontine nuclei, parabrachial, dorsal reticular nuclei, and the external and ventral medial part of the central nucleus of the inferior colliculus. More rostrally at the level of the superior colliculus, terminal fields are found in the medial nucleus of the medial geniculate body, the suprageniculate, pretectal, and mesencephalic reticular nuclei, marking the end of the LAC. In the diencephalon, gracile fibers leave the MLLC and form a crescentlike terminal field along the extreme lateral border of the ventral posterior lateral nucleus (VPL) of the thalamus. Cuneate MLLC fibers terminate in a bandlike formation in the VPL medial to the gracile termination.



The third fiber system, the cuneocerebellar projection, emerges from the cuneate, the external cuneate nuclei, and the “cellular bridge” and immediately enters the ipsilateral inferior cerebellar peduncle. Upon entering the cerebellum, the major fiber component remains ipsilateral and terminates as vertical bands in vermal and paravermal lobules, and lobules I through IVa. The posterior cerebellar lobe contains terminal bands in lobules VII–IX, the copula pyramidis, and the paramedian lobule.



It is concluded that the dorsolateral fiber system conforms to Graybiel's LAC. It is more divergent and probably less modality specific, whereas the medial lemniscal system conforms to the MLLC, which is said to be modality specific, less divergent, and locked to specific sensory-motor response characteristics.



The topography of cerebellar terminal bands indicates that there is sensory-motor representation from all parts of the body to all parts of the cerebellum, at least in the rat.

J. J. Taylor

Papers

Projection systems and terminal localization of dorsal column afferents: An autoradiographic and horseradish peroxidase study in the rat