Vestibular primary afferent projection to the cerebellum of the rabbit

The vestibular primary afferent projection to the cerebellum of the rabbit was studied with retrograde and orthograde tracers. We injected individual lobules of the cerebellum with horseradish peroxidase (HRP) or wheat germ agglutinin-HRP (WGA-HRP). Following these injections the numbers of labeled and unlabeled cells in Scarpa's ganglion were counted. Approximately 64–89% of the cells in Scarpa's ganglion were labeled retrogradely following uvula-nodular injections. About 2% of the cells in the ipsilateral Scarpa's ganglion were labeled after injections of the flocculus. Virtually no cells were labeled following injections of the ventral paraflocculus. The vestibular primary afferent projection to the uvula-nodulus is so extensive that it must be part of a collateral system that also innervates the vestibular nuclei. This collateral projection pattern was confirmed by using fluorescent tracers injected into the uvula-nodulus and vestibular complex. Fluorogold was injected into the uvula-nodulus and peroxidaserhodamine isothiocyanate was injected into the vestibular complex. More than 50% of the neurons in Scarpa's ganglion were double labeled by these subtotal injections. The dense vestibular primary afferent projection to the uvula-nodulus was confirmed by using the C fragment of tetanus toxin (TTC) injected into the labyrinth as an orthograde tracer. With the TTC technique, the vestibular primary afferent projection to the uvula-nodulus terminated exclusively in the ipsilateral granule cell layer of lobules 9d and 10. Much sparser vestibular primary afferent projections were found in the banks of major cerebellar sulci. A barely detectable projection was found to the flocculus and ventral paraflocculus. © 1993 Wiley-Liss, Inc.     

Cholinergic innervation of the cerebellum of rat, rabbit, cat, and monkey as revealed by choline acetyltransferase activity and immunohistochemistry.

The cholinergic innervation of the cerebellar cortex of the rat, rabbit, cat and monkey was studied by immunohistochemical localization of choline acetyltransferase (ChAT) and radiochemical measurement of regional differences in ChAT activity. Four antibodies to ChAT were used to find optimal immunohistochemical localization of this enzyme. These antibodies selectively labeled large mossy fiber rosettes as well as finely beaded terminals with different morphological characterization, laminar distribution within the cerebellar cortex, and regional differences within the cerebellum. Large "grape-like" classic ChAT-positive mossy fiber rosettes that were distributed primarily in the granule cell layer were concentrated, but not exclusively located in three separate regions of the cerebellum in each of the four species studied: 1) The uvula-nodulus (lobules 9 and 10); 2) the flocculus-ventral paraflocculus, and 3) the anterior lobe vermis (lobules 1 and 2). No intrinsic cerebellar neurons were labeled. No cells in either the inferior olive (the origin of cerebellar climbing fibers) or in the locus coeruleus (an origin of noradrenergic fibers) were ChAT-positive. Thin, finely beaded axons, similar to cholinergic axons of the cerebral cortex of the rat, were observed in both the granule cell layer and molecular layer of the cerebellar cortex of the rat, rabbit and cat. The regional differences in ChAT-positive afferent terminations in the cerebellar cortex was for the most part confirmed by regional measurements of ChAT activity in the rat, rabbit, and cat. The three cholinergic afferent projection sites correspond to regions of the cerebellar cortex that receive vestibular primary and secondary afferents. These data imply that a subset of vestibular projections to the cerebellar cortex are cholinergic.     

Secondary vestibular cholinergic projection to the cerebellum of rabbit and rat as revealed by choline acetyltransferase immunohistochemistry, retrograde and orthograde tracers.

Previously we have shown that four regions of the cerebellum, the uvula-nodulus, flocculus, ventral paraflocculus, and anterior lobe 1, receive extensive, but not exclusive, cholinergic mossy fiber projections. In the present experiment we have studied the origin of three of these projections in the rat and rabbit (uvula-nodulus, flocculus, ventral paraflocculus), using choline acetyltransferase (ChAT) immunohistochemistry in combination with a double label, retrogradely transported horseradish peroxidase (HRP). We have demonstrated that in both the rat and rabbit the caudal medial vestibular nucleus (MVN) and to a lesser extent the nucleus prepositus hypoglossus (NPH) contain ChAT-positive neurons. Neurons of the caudal MVN are double-labeled following HRP injections into the uvula-nodulus. HRP injections into the uvula-nodulus also labeled less than 5% of the neurons in the cholinergic vestibular efferent complex. Fewer ChAT-positive neurons in the MVN and some ChAT-positive neurons in the NPH are double-labeled following HRP injections into the flocculus. Almost no ChAT-positive neurons in the MVN and some ChAT-positive neurons in the NPH are double-labeled following HRP injections into the ventral paraflocculus. Injections of Phaseolus leucoagglutinin (PHA-L) into the caudal MVN of both the rat and rabbit demonstrated projection patterns to the uvula-nodulus and flocculus that were qualitatively similar to those observed using ChAT immunohistochemistry. We conclude that the cholinergic mossy fiber pathway to the cerebellum in general and the uvula-nodulus in particular is likely to mediate secondary vestibular information related to postural adjustments.     

Distribution of primary afferent fibers projecting from hindlimb cutaneous nerves to the medulla oblongata in the cat and rat

The dorsal column nuclear complex, one of the most important relays for tactile perception, has well been known to be somatotopically organized. Topographical arrangements of terminal sites of individual cutaneous nerves within the dorsal column nuclei, however, have not been examined systematically, although many studies have been done upon primary afferents to the medulla oblongata, including the dorsal column nuclear complex. Thus, in the present study, distribution of primary afferent fibers projecting from the hindlimb cutaneous nerves to the medulla oblongata was examined in the cat and rat by means of the transganglionic transport method with horseradish peroxidase. Cutaneous primary afferent fibers projecting from the hindlimb to the medulla oblongata were distributed mainly in the ipsilateral gracile nucleus. Terminal labeling in the gracile nucleus was seen at all rostrocaudal levels of the nucleus, occasionally including the nuclear part straddling the midline (the median or accessory nucleus). The labeled axon terminals in the gracile nucleus were more densely distributed in the middle and caudal parts of the nucleus than in the rostral part. Although the fields of termination of the hindlimb cutaneous nerves overlapped in the gracile nucleus, the foci of the terminal labeling of the nerves innervating the distal parts of the hindlimb were located more medially or dorsomedially than those of the nerves innervating the proximal parts. Terminal labeling was further found in a small zone immediately medial to the rostromedial border of the external cuneate nucleus. This hitherto undescribed zone (U zone) contained a small cluster of medium-sized neurons in the cat. Although no particular cell cluster was found in the U zone of the rat, convergence of the primary afferent fibers of the cutaneous nerve from the hindlimb appeared to occur as in the U zone of the cat.     

Peripheral innervation patterns of vestibular nerve afferents in the bullfrog utriculus

Vestibular nerve afferents innervating the bullfrog utriculus differ in their response dynamics and sensitivity to natural stimulation. They also supply hair cells that differ markedly in hair bundle morphology. To examine the peripheral innervation patterns of individual utricular afferents more closely, afferent fibers were labeled by the extracellular injection of horseradish peroxidase (HRP) into the vestibular nerve after sectioning the vestibular nerve medial to Scarpa's ganglion to allow the degeneration of sympathetic and efferent fibers. The peripheral arborizations of individual afferents were then correlated with the diameters of their parent axons, the regions of the macula they innervate, and the number and type of hair cells they supply. The utriculus is divided by the striola, a narrow zone of distinctive morphology, into medial and lateral parts. Utricular afferents were classified as striolar or extrastriolar according to the epithelial entrance of their parent axons and the location of their terminal fields. In general, striolar afferents had thicker parent axons, fewer subepithelial bifurcations, larger terminal fields, and more synaptic endings than afferents in extrastriolar regions. Afferents in a juxtastriolar zone, immediately adjacent to the medial striola, had innervation patterns transitional between those in the striola and more peripheral parts of the medial extrastriola. Most afferents innervated only a single macular zone. The terminal fields of striolar afferents, with the notable exception of a few afferents with thin parent axons, were generally confined to one side of the striola. Hair cells in the bullfrog utriculus have previously been classified into four types based on hair bundle morphology (Lewis and Li: Brain Res. 83:35–50, 1975). Afferents in the extrastriolar and juxtastriolar zones largely or exclusively innervated Type B hair cells, the predominant hair cell type in the utricular macula. Striolar afferents supplied a mixture of four hair cell types, but largely contacted Type B and Type C hair cells, particularly on the outer rows of the medial striola. Afferents supplying more central striolar regions innervated fewer Type B and larger numbers of Type E and Type F hair cells. Striolar afferents with thin parent axons largely supplied Type E hair cells with bulbed kinocilia in the innermost striolar rows. © 1994 Wiley-Liss, Inc.     

Physiological and anatomical characteristics of primary vestibular afferent neurons in the bullfrog

Intracellular recordings were made in the VIIIth nerve of the bullfrog (Rana catesbiana) to measure the membrane characteristics and obtain records of spontaneous and evoked spike activity of primary semicircular canal afferents. Physiological stimulation of the canals was achieved by rotating the preparation on a servomotor driven turntable with the animals' head centered in the rotational axis. The responses of each neuron to sinusoidal rotations at frequencies of 0.05Hz, 0.5Hz and for impulsive accelerations of 400 deg/sec2 were obtained. Membrane characteristics measured included the cell resting and action potential amplitude, and spike-activation threshold for applied currents. Physiologically characterized neurons were injected with horseradish peroxidase by applying pneumatic pressure and/or iontophoretic currents to the micropipettes containing 5% HRP in 1 M KCI. Following survival times of 12--48 h, the VIIIth nerve and attached vestibular end organ was removed for histochemical processing using a diaminobenzidine procedure to visualize the HRP reaction product. Light microscopy was used to discern the anatomical features of the neurons and to trace their peripheral dendritic trajectories from the ganglion to their termination(s) in the crista. Our studies have revealed that the bullfrog's primary vestibular afferents are characterized by a broad range of soma and axon diameters which correspond to an equally broad range of spontaneous and evoked activity characteristics. The largest neurons had more irregular spontaneous firing rates and consistently exhibited the greatest gain and smallest phase shifts with respect to head acceleration. These neurons consistently terminated at or near the central region of the crista. On the other hand, the smallest neurons were characterized by having the most regular spontaneous discharge patterns, the lowest gains, and greatest phase shifts with respect to head acceleration. Our findings are thus consistent with the view that the anatomical features of the primary vestibular neurons are important in determining the neuron's physiological characteristics. In terms of response dynamics our observations indicate that the receptors in the frog's crista ampullaris are heterogeneous and differentially sensitive to a wide range of stimulus frequencies.     

Direct connections of primary afferent fibers with spinal neurons projecting to the facial nucleus in the cat

Spinal neurons projecting to the facial nucleus, located in the lateral portion of lamina V from the C1 to C4 segments, had long dendrites extending into lamina IV. The detailed morphology of these spinal neurons was confirmed using the Golgi impregnation method. Dorsal root fibers were distributed mainly to lamina IV at the C1 and C2 segments. These findings indicated the possibility of direct connections between the primary afferent fibers and spinal neurons projecting to the facial nucleus in lamina IV of the upper cervical cords. In order to confirm the connections, electron microscopic observation was carried out on the dorsal horn after injections of horseradish peroxidase into the facial nucleus and dorsal root fibers.     

Characterization of the primary spinal afferent innervation of the mouse colon using retrograde labelling

Visceral pain is the most common form of pain produced by disease and is thus of interest in the study of gastrointestinal (GI) complaints such as irritable bowel syndrome, in which sensory signals perceived as GI pain travel in extrinsic afferent neurones with cell bodies in the dorsal root ganglia (DRG). The DRG from which the primary spinal afferent innervation of the mouse descending colon arises are not well defined. This study has combined retrograde labelling and immunohistochemistry to identify and characterize these neurones. Small to medium-sized retrogradely labelled cell bodies were found in the DRG at levels T8-L1 and L6-S1. Calcitonin gene-related peptide (CGRP)- and P2X3-like immunoreactivity (LI) was seen in 81 and 32%, respectively, of retrogradely labelled cells, and 20% bound the Griffonia simplicifolia-derived isolectin IB4. CGRP-LI and IB4 were co-localized in 22% of retrogradely labelled cells, whilst P2X3-LI and IB4 were co-localized in 7% (vs 34% seen in the whole DRG population). Eighty-two per cent of retrogradely labelled cells exhibited vanilloid receptor 1-like immunoreactivity (VR1-LI). These data suggest that mouse colonic spinal primary afferent neurones are mostly peptidergic CGRP-containing, VR1-LI, C fibre afferents. In contrast to the general DRG population, a subset of neurones exist that are P2X3 receptor-LI but do not bind IB4.     

Direct connections of primary afferent fibers with central cervical nucleus neurons projecting to the cerebellum in the cat

The synaptic organization of the central cervical nucleus (CCN) in the cat was examined by means of anterograde and retrograde axonal transport of horseradish peroxidase and the anterograde degeneration tracing technique. Light microscopic studies confirmed that cervical primary afferents terminate heavily in the region of CCN cerebellar projection neurons and electron microscopic findings demonstrated that primary afferent fibers of cervical dorsal roots form synapses mainly with distal dendrites of CCN neurons projecting to the cerebellum.     

Response patterns of primary vestibular neurons to thermal and rotational stimuli

The neural responses of vestibular ganglion cells to quantified thermal and rotational stimulation were studied in anesthetized squirrel monkeys. All cells exhibited a spontaneous discharge rate with the average being 72 ± 27.8 spikes/sec. The observed increases and decreases in discharge rates during sinusoidal angular rotation were consistent with the morphological polarization patterns of the hair cells. Likewise, the ganglion cell responses resulting from thermal stimulation by a controlled temperature air stream directed on the lateral portion of the horizontal canal were in agreement with those expected on the basis of Barany's convective current theory and the polarization pattern of the hair cells.In an attempt to determine the relationship between thermal and rotational stimuli, the sensitivities of individual cells to both types of stimuli were determined. Those cells most sensitive to angular rotation were also most sensitive to caloric stimulation. An equivalency value for the two types of stimuli was defined and found to be 1.84 °C/radian·sec^?1.The global response characteristics of the vestibular endorgan were studied by simultaneous thermal and rotational stimuli. The thermal stimulus was used to bias the cupula at a new equilibrium position while a rotational stimulus was used to determine the differential sensitivity at that cupular position. For the cells with a more regular spontaneous discharge rate, the differential sensitivity was observed to be a function of the instantaneous discharge rate. No such relationship was observed for the more irregular cells.     

Patterns of primary afferent termination in the external cuneate nucleus from cervical axial muscles in the cat

Using the method of transganglionic transport of horseradish peroxidase (HRP), the distribution of primary afferent projections was examined in the external cuneate nucleus (ECN) from different muscle groups in the forequarter of the cat. The terminal zones of afferent fibers from three shoulder muscles--clavotrapezius, acromiotrapezius, and spinotrapezius--were compared to projections from suboccipital muscles, dorsal neck extensors, and muscles of the proximal forelimb. Each muscle group had a labelled terminal zone that occupied a different subvolume of the ECN. The zone labelled from trapezius muscles formed a continuous column in the ECN running from the caudal pole of the nucleus to a level 3.0 mm rostral to the obex. Terminal zones of suboccipital muscles and dorsal neck extensors formed longer columns that extended into the most rostral tip of the ECN, while those of proximal forelimb muscles formed shorter columns confined to the caudal two-thirds of the ECN. At comparable cross-sectional levels in the caudal and middle portions of the ECN, terminal zones from proximal limb muscles were located most dorsomedially, while those from shoulder muscles, dorsal neck muscles, and suboccipital muscles were located in progressively more ventral and lateral regions. The subvolume of the ECN occupied by projections from cervical axial muscles was estimated to be more than 40% of the volume of the nucleus, suggesting that the ECN has a major role in the transmission of sensory information from axial musculature to the cerebellum. Following exposure of all muscle nerves to tracer, a second labelled zone was also identified close to the ECN in the descending vestibular nucleus at transverse levels 2.0-3.0 mm rostral to the obex. Here, reaction product was concentrated around a circumscribed collection of medium-sized, multipolar cells.     

Topographical organization of subicular neurons projecting to subcortical regions

Direct projections from the subiculum to the septum, thalamus, and hypothalamus were studied in the rat by the fluorescent retrograde double-labeling technique with Fast blue and Diamidino yellow. The results confirm and extend the previously reported findings. The dorsal subiculum projects primarily to the lateral septum, anterior and midline thalamus, and mammillary complex. The distribution areas of cell bodies of these projection neurons are substantially segregated, depending on their target region, and few single neurons project to two of the target regions by way of axon collaterals. The ventral subiculum projects mainly to the lateral septum, midline thalamus, and ventromedial hypothalamic area. The distribution areas of cell bodies of these projection neurons are considerably overlapped with one another, and a number of single neurons send axon collaterals to two of the lateral septum, midline thalamus, and ventromedial hypothalamic area. It is, thus, indicated that the populations of subicular neurons projecting to each of the subcortical structures examined are more distinctly segregated in the dorsal subiculum than in the ventral subiculum.     

Distribution of the vestibular neurons projecting to the oculomotor and trochlear nuclei in rabbits

Horseradish peroxidase and Fast Blue were injected into the oculomotor and trochlear nuclei of rabbits so as to study the distribution of vestibular neurons that project to these nuclei. After the oculomotor nucleus was injected, labelled neurons were found in the superior, medial, and descending vestibular nuclei as well as in cell group Y. In the superior nucleus, most of the neurons (510 +/- 46) were ipsilateral to the injection, although contralaterally labelled neurons were also observed (104 +/- 19) more peripherally. In cell group Y, 186 +/- 24 contralaterally labelled neurons were observed, whereas hardly any (8 +/- 3) were found on the ipsilateral side. The largest group of labelled neurons (811 +/- 65) constituted a neuronal band located contralaterally in the medial nucleus and rostral part of the descending nucleus. This band rostromedially continued with the caudal part of the group of internuclear neurons of the abducens nucleus. Only 190 +/- 31 neurons were labelled in the medial and descending nucleus ipsilateral to the injected oculomotor nucleus. After injection of the trochlear nucleus, labelled neurons were found in the ipsilateral superior nucleus and contralateral medial and descending nuclei: a few labelled cells were also observed in the ipsilateral medial and descending nuclei as well as in the contralateral cell group Y.     

Effects of cortical and thalamic lesions upon primary afferent terminations, distributions of projection neurons, and the cytochrome oxidase pattern in the trigeminal brainstem complex.

Early postnatal lesions of the primary somatosensory cortex alter the vibrissa-related cytochrome oxidase (CO) pattern in nucleus principalis (PrV) of the rat's trigeminal (V) brainstem complex (Erzurumlu and Ebner, '88: Dev. Brain Res. 44:302-308). At present, the reason for this change is not clear. It may be that the corticotrigeminal projection is necessary for the maintenance of vibrissa-related patterns in PrV. However, it is also possible that the loss of the normal pattern of CO activity reflects a change in the organization of brainstem cells resulting from transneuronal retrograde degeneration. In order to address this question, we made lesions of either the primary somatosensory cortex (S-I) or ventrobasal thalamus (VB) in newborn rats and directly assayed distribution of V primary afferents by transganglionic transport of horseradish peroxidase and V-thalamic neurons by retrograde transport of either fluorogold or true blue. Neonatal cortical and thalamic lesions produced no qualitative change in the distribution of primary afferent terminals in either PrV or V subnucleus interpolaris (SpI) beyond that which could be attributed to shrinkage of the brainstem resulting from retrograde degeneration. Most importantly, the "patchy" pattern of terminations observed in normal rats remained apparent in the brain-damaged animals. The normal distribution of V-thalamic neurons in PrV was disrupted by both cortical and thalamic lesions. These cells are normally patterned in a way that matches the distribution of primary afferent terminals and thus that of the mystacial vibrissae. This was not the case in the neonatally brain-damaged rats. Taken together, these results are consistent with the conclusion that neonatal cortical and thalamic lesions disrupt the normal CO pattern in PrV primarily because of their effects upon the patterning of brainstem cells. The present findings demonstrate further that clustering of primary afferents does not require a normal complement of postsynaptic neurons.     

Thalamic innervation of striatal and subthalamic neurons projecting to the rat entopeduncular nucleus

The present study analyses the anatomical arrangement of the projections linking the Wistar rat parafascicular thalamic nucleus (PF) and basal ganglia structures, such as the striatum and the subthalamic nucleus (STN), by using neuroanatomical tract-tracing techniques. Both the thalamostriatal and the striato-entopeduncular projections were topographically organized, and several areas of overlap between identified circuits were noticed, sustaining the existence of up to three separated channels within the Nauta-Mehler loop. Thalamic afferents arising from dorsolateral PF territories are in register with striatofugal neurons located in dorsolateral striatal areas, which in turn project to dorsolateral regions of the entopeduncular nucleus (ENT). Medial ENT regions are innervated by striatal neurons located within medial striatal territories, these neurons being the target for thalamic afferents coming from medial PF areas. Finally, afferents from neurons located in ventrolateral PF areas approached striatal neurons in ventral and lateral striatal territories, which in turn project towards ventral and lateral ENT regions. Efferent STN neurons projecting to ENT were found to be the apparent postsynaptic target for thalamo-subthalamic axons. The thalamo-subthalamic projection was also topographically organized. Medial, central and lateral STN territories are innervated by thalamic neurons located within medial, ventrolateral and dorsolateral PF areas, respectively. Thus, each individual PF subregion projects in a segregated fashion to specific parts of the striato-entopeduncular and subthalamo-entopeduncular systems. These circuits enabled the caudal intralaminar nuclei to modulate basal ganglia output.     

Cervical primary afferent input to vestibulospinal neurons projecting to the cervical dorsal horn: An anterograde and retrograde tracing study in the cat

Vestibulospinal neurons in the caudal half of the medial and descending vestibular nuclei terminate in the cervical spinal cord, not only in the ventral horn and intermediate zone but also in the dorsal horn. The purpose of the present study was to examine whether the areas containing these vestibulospinal neurons are reached by cervical primary afferents. In one group of experiments, wheat germ agglutinin-horseradish peroxidase conjugate and horseradish peroxidase were pressure injected into spinal ganglia C₂-C₈ and revealed anterogradely labeled fibers and boutons in the caudal part (caudal to the dorsal cochlear nucleus) of the ipsilateral medial and descending vestibular nuclei. This projection was verified in experiments in which wheat germ agglutinin-horseradish peroxidase conjugate was microiontophoretically injected into the caudal half of either the medial or the descending vestibular nuclei and revealed retrogradely labeled cells only in ipsilateral spina ganglia C₂-C₇, with a maximum of cells in C₃. In another group of experiments, after microiontophoretic injections of Phaseolus vulgaris leucoagglutinin or Biocytin into either the medial or the descending vestibular nuclei, anterogradely labeled fibers and boutons were present in the cervical spinal cord, mainly bilaterally in the dorsal horn (laminae I–VI) but also, to a lesser extent, in the ventral horn and intermediate zone. The existence of a loop that relays cervical primary afferent information to vestibulospinal neurons projecting to the cervical spinal cord, in particular the dorsal horn, may have implications for vestibular control over local information processing in the cervical dorsal horn. © 1995 Wiley-Liss, Inc.     

Substance P afferent terminals innervate vagal preganglionic neurons projecting to the trachea of the ferret

Airway disorders, such as asthma and chronic obstructive bronchitis, are, in part, due to abnormalities in the nervous control of the airways. However, the ultrastructural circuitry and neurochemical anatomy of afferents modulating the output of airway-related vagal preganglionic neurons (VPNs) in the nucleus ambiguus are poorly understood. We have examined the potential role of substance P (SP) immunoreactive afferents in the regulation of anatomically identified airway VPNs. Cholera toxin b-subunit conjugated to horseradish peroxidase was used as a retrograde cell body tracer to identify the central VPNs innervating the extra-thoracic trachea. Immunocytochemistry was employed to identify SP afferents. The external formation of the nucleus ambiguus was examined by electron microscopy using a simultaneous double labeling method. Cell bodies of tracheal VPNs were 31.7 +/- 1.18 x 23.0 +/- 1.3 microm (means +/- S.E.M.) in size, contained abundant endoplasmic reticulum, had a round nucleus with a prominent nucleolus, no satellite body and displayed somatic and dendritic spines. Somato-somatic appositions, somato-dendritic appositions without intervening glial processes and dendritic "bundling" commonly seen in esophageal motoneurons were not observed. The ultrastructural morphology of tracheal VPNs were also clearly distinguishable from pharyngeal and laryngeal motoneurons in other divisions of the nucleus ambiguus which lack somatic spines. These data are consistent with the hypothesis that differences in the ultrastructure and synaptology of the different divisions of the nucleus ambiguus may be associated with specific physiological functions. The mean size (+/- S.E.M.) of SP nerve terminals was 1.57 +/- 0.06 x 0.79 +/- 0.03 microm. SP terminals formed 17.5% of the axo-dendritic and 15.9% of the axo-somatic synapses which were observed upon retrogradely labeled tracheal VPNs. Synaptic contacts observed were both symmetric and asymmetric. These synaptic interactions define, in part, the neurochemical anatomy of neuronal circuits modulating vagal preganglionic control of tracheal functions.     

Somatotopic organization of the primary sensory trigeminal neurons in the hagfish, Eptatretus burgeri

Primary sensory trigeminal projections were investigated in the hagfish following application of horseradish peroxidase (HRP) to the sensory branches. In our control preparations we were able to distinguish five sensory ganglia and their respective nerves. HRP application confirmed the almost exclusive relation of each of these nerves to their respective ganglia, with very little overlap. In normal frontal sections of the medulla oblongata, five columns of fibers surrounded by neuronal cell bodies could be clearly distinguished, but the number is probably fortuitous, for there was no one-on-one relationship with the five trigeminal ganglia. From their peripheral connections, we surmised that columns 1 and 3 handle general cutaneous sensation, columns 2, 4, and 5 handle taste sensation, and column 5 handles general mucous cutaneous sensation conveyed by utricular ganglion cells. Dorsally located columns received projections from nerves with dorsal peripheral connections, and more ventrally located columns received projections from nerves with ventral peripheral connections. This relation is the reverse of that seen in other vertebrates.     

Cholinergic innervation of principal neurons in the cochlear nucleus of the Mongolian gerbil

Principal neurons in the ventral cochlear nucleus (VCN) receive powerful ascending excitation and pass on the auditory information with exquisite temporal fidelity. Despite being dominated by ascending inputs, the VCN also receives descending cholinergic connections from olivocochlear neurons and from higher regions in the pontomesencephalic tegmentum. In Mongolian gerbils, acetylcholine acts as an excitatory and modulatory neurotransmitter on VCN neurons, but the anatomical structure of cholinergic innervation of gerbil VCN is not well described. We applied fluorescent immunohistochemical staining to elucidate the development and the cellular localization of presynaptic and postsynaptic components of the cholinergic system in the VCN of the Mongolian gerbil. We found that cholinergic fibers (stained with antibodies against the vesicular acetylcholine transporter) were present before hearing onset at P5, but innervation density increased in animals after P10. Early in development cholinergic fibers invaded the VCN from the medial side, spread along the perimeter and finally innervated all parts of the nucleus only after the onset of hearing. Cholinergic fibers ran in a rostro‐caudal direction within the nucleus and formed en‐passant swellings in the neuropil between principal neurons. Nicotinic and muscarinic receptors were expressed differentially in the VCN, with nicotinic receptors being mostly expressed in dendritic areas while muscarinic receptors were located predominantly in somatic membranes. These anatomical data support physiological indications that cholinergic innervation plays a role in modulating information processing in the cochlear nucleus.     

Monosynaptic vestibular nerve input to vestibular and adjacent reticular neurons projecting into the ascending medial longitudinal fasciculus.

This electrophysiological study provides evidence that cat neurons both within and ventral to the cytoarchitectural boundary of the vestibular nuclei project into the region of the contralateral ascending medial longitudinal fasciculus. Many of these neurons, including the more ventral ones, receive monosynaptic input from the vestibular nerve. The vestibular complex thus appears to include part of what traditionally has been called reticular formation.