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Neuronal architecture of the dorsal nucleus (cochlear nucleus) of the frog, Rana pipiens pipiens.

The neuronal architecture of the dorsal nucleus of the Northern leopard frog (Rana pipiens pipiens), which is a homolog of the cochlear nucleus of mammals and birds, was investigated. Our study showed that the frog dorsal nucleus contains a number of morphologically distinct cell types that are discernible in terms of the cellular architecture as derived from Nissl-stained material and in terms of the dendritic profile as revealed by horseradish peroxidase-filled single neurons. These cell types are bushy cells, bipolar (or fusiform) cells, octopus cells, stellate cells, giant cells, radiate (or round) cells, and a variety of small cells. The different cell types occupy different regions of the nucleus. Therefore, our results suggest that the dorsal nucleus should no longer be considered to be a uniform nucleus containing a homogeneous population of neurons. Homologies of these cell types with those described in other vertebrate species, including mammals, are proposed.

Animals↗

Synapses formed by olivocochlear axon branches in the mouse cochlear nucleus.

Cochlear nucleus branches of thick olivocochlear axons were labeled by injections of horseradish peroxidase into the spiral ganglion of the cochlear basal turn in mice. Six labeled axons were traced by light microscopy, and selected portions of seven branches were sectioned serially for electron microscopic examination. Axonal branches most frequently terminated near certain granule cell regions of the ventral cochlear nucleus. This article describes terminals, synapses, and postsynaptic elements of these olivocochlear branches. The olivocochlear branches had both terminal and en passant boutons that contained round vesicles and made asymmetric synapses with other neuronal processes. About a quarter of the synapses also possessed additional specializations, postsynaptic, or subjunctional bodies. Mossy terminals, a multisynaptic type of terminal commonly found in granule cell regions, were not found arising from any of the labeled branches. No somatic synapses were found, although contacts with cell bodies were occasionally observed. The predominant synaptic target of olivocochlear branches were what appeared to be dendrites of large diameter. At least some of these large dendrites received multiple synapses from a single labeled olivocochlear branch. The morphological characteristics of reconstructed dendrites suggest that multipolar cells might be predominant targets for the medial olivocochlear system in the cochlear nucleus. This was demonstrated in one case in which a large dendrite was followed to its cell body of origin.

Animals↗

Projections from the ventral cochlear nucleus to the dorsal cochlear nucleus in rats.

Local circuit interactions between the dorsal and ventral divisions of the cochlear nucleus are known to influence the evoked responses of the resident neurons to sound. In the present study, we examined the projections of neurons in the ventral cochlear nucleus to the dorsal cochlear nucleus by using retrograde transport of biotinylated dextran amine injected into restricted but different regions of the dorsal cochlear nucleus. In all cases, we found retrogradely labeled granule, unipolar brush, and chestnut cells in the granule cell domain, and retrogradely labeled multipolar cells in the magnocellular core of the ventral cochlear nucleus. A small number of the labeled multipolar cells were found along the margins of the ventral cochlear nucleus, usually near the boundaries of the granule cell domain. Spherical bushy, globular bushy, and octopus cells were not labeled. Retrogradely-labeled auditory nerve fibers and the majority of labeled multipolar neurons formed a narrow sheet extending across the medial-to-lateral extent of the ventral cochlear nucleus whose dorsoventral position was topographically related to the injection site. Labeled multipolar cells within the core of the ventral cochlear nucleus could be divided into at least two distinct groups. Planar neurons were most numerous, their somata found within the associated band of labeled fibers, and their dendrites oriented within this band. This arrangement mimics the organization of isofrequency contours and implies that planar neurons respond best to a narrow range of frequencies. In contrast, radiate neurons were infrequent, found scattered throughout the ventral cochlear nucleus, and had long dendrites oriented perpendicular to the isofrequency contours. This dendritic orientation suggests that radiate neurons are sensitive to a broad range of frequencies. These structural differences between planar and radiate neurons suggest that they subserve separate functions in acoustic processing.

Animals↗

Unit responses in ventral cochlear nucleus reflect cochlear coding of rapid frequency sweeps.

This study examines the encoding of rapid frequency sweeps in single units of the ventral cochlear nucleus (VCN). Sweeps were designed to explore the role of cochlear mechanics in shaping the temporal responses across cells in the VCN. The time course of frequency change for rapidly rising frequency sweeps theoretically produced simultaneous displacement maxima by cancelling travel time along the cochlear partition. Rising sweeps with longer time courses only partially canceled travel time, while falling sweeps had time courses of frequency change equal to or greater than travel time. Falling sweeps thus augmented normal travel time. Latency of unit firing to sweeps across unit characteristic frequency (CF) reflected cochlear delay-line mechanics. The latency-CF functions agreed with predictions from travel-time estimates for rising-frequency sweeps, but responses to falling sweeps were less predictable.

Acoustic Stimulation↗

Multipolar cells in the ventral cochlear nucleus project to the dorsal cochlear nucleus and the inferior colliculus.

When retrograde markers are placed in the dorsal cochlear nucleus two classes of labeled cells are found in the ventral cochlear nucleus. These are multipolar cells and granule cells. The structure and distribution of labeled multipolar cells greatly resemble those seen following injection of retrograde markers into the contralateral inferior colliculus. When one retrograde marker is placed in the dorsal cochlear nucleus and another simultaneously placed into the contralateral inferior colliculus, large numbers of multipolar cells containing both markers are found in the ventral cochlear nucleus. These findings show that all or most cells in the ventral cochlear nucleus that project to the inferior colliculus also send collaterals to the ipsilateral dorsal cochlear nucleus.

Animals↗

Mossy fiber projections from the cuneate nucleus to the cochlear nucleus in the rat.

A reciprocal connection is known to exist between the cuneate nucleus, which is a first-order somatosensory nucleus, and the cochlear nucleus, which is a first-order auditory nucleus. We continued this line of study by investigating the fiber endings of this projection in the cochlear nucleus of rats using the neuronal tracer Phaseolus vulgaris leucoagglutinin in combination with ultrastructural and immunocytochemical analyses. In the cochlear nucleus, mossy fiber terminals had been described and named for their morphologic similarity to those in the cerebellum, but their origins had not been discovered. In the present study, we determined that the axonal projections from the cuneate region gave rise to mossy fiber terminals in the granule cell regions of the ipsilateral cochlear nucleus. The cuneate mossy fibers appear to be excitatory in nature, because they are filled with round synaptic vesicles, they make asymmetric synapses with postsynaptic targets, and they are labeled with an antibody to glutamate. The postsynaptic targets of the mossy fibers include dendrites of granule cells. This projection onto the granule cell interneuron circuit of the cochlear nucleus indicates that somatosensory cues are intimately involved with information processing at this early stage of the auditory system.

Animals↗

Glycine immunoreactivity of multipolar neurons in the ventral cochlear nucleus which project to the dorsal cochlear nucleus.

Certain distinct populations of neurons in the dorsal cochlear nucleus are inhibited by a neural source that is responsive to a wide range of acoustic frequencies. In this study, we examined the glycine immunoreactivity of two types of ventral cochlear nucleus neurons (planar and radiate) in the rat which project to the dorsal cochlear nucleus (DCN) and thus, might be responsible for this inhibition. Previously, we proposed that planar neurons provided a tonotopic and narrowly tuned input to the DCN, whereas radiate neurons provided a broadly tuned input and thus, were strong candidates as the source of broadband inhibition (Doucet and Ryugo [1997] J. Comp. Neurol. 385:245-264). We tested this idea by combining retrograde labeling and glycine immunohistochemical protocols. Planar and radiate neurons were first retrogradely labeled by injecting biotinylated dextran amine into a restricted region of the dorsal cochlear nucleus. The labeled cells were visualized using streptavidin conjugated to indocarbocyanine (Cy3), a fluorescent marker. Sections that contained planar or radiate neurons were then processed for glycine immunocytochemistry using diaminobenzidine as the chromogen. Immunostaining of planar neurons was light, comparable to that of excitatory neurons (pyramidal neurons in the DCN), whereas immunostaining of radiate neurons was dark, comparable to that of glycinergic neurons (cartwheel cells in the dorsal cochlear nucleus and principal cells in the medial nucleus of the trapezoid body). These results are consistent with the hypothesis that radiate neurons in the ventral cochlear nucleus subserve the wideband inhibition observed in the dorsal cochlear nucleus.

Animals↗

Fine structure and neurotransmitter cytochemistry of neurons in the rat ventral cochlear nucleus projecting to the ipsilateral dorsal cochlear nucleus.

The neural tracer wheat germ agglutinin conjugated to horse radish peroxidase was injected into the rat dorsal cochlear nucleus and acoustic stria. Some labelled neurons in the ipsilateral ventral cochlear nucleus were found as a result. These neurons were studied at the ultrastructural level, and their axo-somatic synaptic profile and glycine immunoreactivity were determined. Most neurons were glycine negative and classified as type I multipolar neurons. The latter showed a different synaptic profile from that of neurons projecting to the contralateral inferior colliculus or cochlear nucleus. This suggests the presence of differing populations of multipolar cells based on their synaptic profile. Few labelled multipolar neurons of type II were found, which appeared glycine negative and, rarely, glycine positive. The latter show an ultrastructure and axo-somatic profile similar to that of glycinergic commissural neurons in the dorsal and ventral cochlear nucleus. In particular, about one-third of boutons contained round synaptic vesicles, which are believed to contain an excitatory neurotransmitter. The ultrastructural analysis of the synaptic boutons in the cochlear nucleus confirms the presence of numerous cases of colocalization of glycine and GABA where flat and pleomorphic synaptic vesicles are mixed. The present study is in accordance with previous tract-tracing light microscopic studies which have indicated that large glycinergic neurons in the ventral cochlear nucleus act as broad-band inhibitory neurons in microcircuits of the dorsal cochlear nucleus and contralateral cochlear nucleus.

Animals↗

Serotonin projection patterns to the cochlear nucleus.

The cochlear nucleus is well known as an obligatory relay center for primary auditory nerve fibers. Perhaps not so well known is the neural input to the cochlear nucleus from cells containing serotonin that reside near the midline in the midbrain raphe region. Although the specific locations of the main, if not sole, sources of serotonin within the dorsal cochlear nucleus subdivision are known to be the dorsal and median raphe nuclei, sources of serotonin located within other cochlear nucleus subdivisions are not currently known. Anterograde tract tracing was used to label fibers originating from the dorsal and median raphe nuclei while fluorescence immunohistochemistry was used to simultaneously label specific serotonin fibers in cat. Biotinylated dextran amine was injected into the dorsal and median raphe nuclei and was visualized with Texas Red, while serotonin was visualized with fluorescein. Thus, double-labeled fibers were unequivocally identified as serotoninergic and originating from one of the labeled neurons within the dorsal and median raphe nuclei. Double-labeled fiber segments, typically of fine caliber with oval varicosities, were observed in many areas of the cochlear nucleus. They were found in the molecular layer of the dorsal cochlear nucleus, in the small cell cap region, and in the granule cell and external regions of the cochlear nuclei, bilaterally, of all cats. However, the density of these double-labeled fiber segments varied considerably depending upon the exact region in which they were found. Fiber segments were most dense in the dorsal cochlear nucleus (especially in the molecular layer) and the large spherical cell area of the anteroventral cochlear nucleus; they were moderately dense in the small cell cap region; and fiber segments were least dense in the octopus and multipolar cell regions of the posteroventral cochlear nucleus. Because of the presence of labeled fiber segments in subdivisions of the cochlear nucleus other than the dorsal cochlear nucleus, we concluded that the serotoninergic projection pattern to the cochlear nucleus is divergent and non-specific. Double-labeled fiber segments were also present, but sparse, in the superior olive, localized mainly in periolivary regions; this indicated that the divergence of dorsal and median raphe neurons that extends throughout regions of the cochlear nucleus also extended well beyond the cochlear nucleus to include at least the superior olivary complex as well.

Afferent Pathways↗

Projections from the ventral cochlear nucleus to the inferior colliculus and the contralateral cochlear nucleus in guinea pigs.

Multipolar cells in the ventral cochlear nucleus are the source of projections to numerous brainstem auditory nuclei, including the contralateral and ipsilateral inferior colliculi and the contralateral cochlear nucleus. Multiple fluorescent tracers were used to label the multipolar cells that project to each of these targets. Following injections of different tracers into each target, the ventral cochlear nucleus was examined for the presence of cells that contained more than one tracer. Such cells were never observed. In contrast, double-labeled cells were common in the dorsal cochlear nucleus, where cells frequently contained the two tracers that were injected into the ipsilateral and contralateral inferior colliculi. The distribution and somatic morphology of cells in the ventral cochlear nucleus that project to each of the three targets were examined. Each population contained cells with somas that ranged in shape from elongated to rounded, but there were differences in soma size. Projections to the ipsilateral and contralateral inferior colliculi arise predominantly from small to medium-sized cells, the average size being slightly less for cells with projections to the ipsilateral colliculus. Projections to the contralateral cochlear nucleus arise from cells with somas that range in size from small to large, including cells much larger than those that projected to either inferior colliculus. On the basis of these results, we conclude that projections from the ventral cochlear nucleus to the ipsilateral and contralateral inferior colliculi and to the contralateral cochlear nucleus arise in three different populations of multipolar cells.

Analysis of Variance↗

The postnatal growth of cochlear nucleus subdivisions and neuronal somata of the anteroventral cochlear nucleus in the Mongolian gerbil (Meriones unguiculatus).

We have analysed the postnatal volume increase of the cochlear nucleus subdivisions from birth to senescence in the Mongolian gerbil. All subdivisions showed the most prominent growth in the 2nd week of life before the onset of hearing on day 12. After the onset of hearing, the dorsal and the postero-ventral portion of the cochlear nucleus showed only a moderate degree of further growth. The antero-ventral cochlear nucleus, however, almost doubled in volume after the onset of hearing, reaching a maximum in 4-month-old animals. In ageing gerbils, we observed a small but systematic reduction of the cochlear nucleus volume. To further assess which elements contribute to the growth of the antero-ventral cochlear nucleus after the onset of hearing, we analysed the somal size of the neurones. Before the onset of hearing, somal and volume growth were highly correlated. However, somata had reached an adult size by the onset of hearing. We conclude that the growth of the antero-ventral cochlear nucleus after the onset of hearing is due to changes in the neuropil.

Age Factors↗

Development and degeneration of hearing in the C57/b16 mouse: relation of electrophysiologic responses from the round window and cochlear nucleus to cochlear anatomy and behavioral responses.

In disorders of the auditory organ associated with hearing loss, a correlation of the anatomical, behavioral and electrophysiological studies furnishes the investigator several parameters of study to better comprehend the hearing mechanism and the various anatomical structural dysfunctions that relate to the hearing loss. Mice are interesting to study because they carry several types of genes for hereditary deafness. Until recently, it was very difficult to train mice for behavioral hearing threshold testing; however, having developed such a method, the next step was to study the normal mice in these three parameters and extend the studies to mice with hereditary hearing loss. The type of mouse studied in this research was the C57/b16 strain which carries such a gene. The results of the studies indicate that the results of the behavioral and electrophysiological studies relate well to each other, and that light microscopic studies (if taken alone) do not furnish us with the actual functional capability of the structures of the inner ear. It is recommended (and it would be most valuable) that efforts should be made to collect and study human temporal bones with all three parameters of study for a better understanding of the structures of the human inner ear. Reference is made and recommendations given as to the parameters of studies while performing human electrocochleography.

Acoustic Stimulation↗

Uptake and release of gamma-aminobutyric acid in the guinea pig cochlear nucleus after axotomy of cochlear and centrifugal fibers.

This study attempts to determine if gamma-aminobutyric acid (GABA) may be a transmitter of cochlear nerve fibers projecting from the cochlea to the cochlear nucleus, and of centrifugal fibers projecting to the cochlear nucleus via the trapezoid body and the acoustic striae of the medulla. The uptake and the electrically evoked release of exogenous [14C]GABA were measured, in vitro, in the three major subdivisions of the guinea pig cochlear nucleus; the anteroventral, posteroventral, and dorsal cochlear nuclei. These activities were compared using unlesioned animals, animals with bilateral cochlear ablations, and animals whose trapezoid body and acoustic striae were interrupted on the right side of the medulla. Subdivisions from unlesioned animals took up [14C]GABA, achieving concentrations in the tissues that were 11-19 times that in the medium. Electrical stimulation evoked a Ca2+-dependent release of [14C]GABA from each subdivision. Bilateral cochlear ablation, which presumably destroyed the cochlear nerve fibers, had no effect on [14C]GABA uptake and release. Section of the trapezoid body and the acoustic striae on the right side of the medulla typically severed all known connections of the right posteroventral and dorsal cochlear nuclei with the rest of the brain, but left intact many connections involved with the right anteroventral cochlear nucleus. This lesion partially depressed [14C]GABA uptake and release in the right posteroventral and dorsal cochlear nuclei, but not in the right anteroventral cochlear nucleus. These findings suggest that one or more of the centrifugal tracts projecting to the cochlear nucleus may be GABAergic, 88% or more of the cochlear nerve fibers probably are not GABAergic, and some neurons of the cochlear nucleus are probably GABAergic.

Animals↗

Synaptophysin immunoreactivity in the cochlear nucleus after unilateral cochlear or ossicular removal.

This study determined if unilateral cochlear removal in adult guinea pigs led to synaptic loss followed by synaptogenesis in the cochlear nucleus (CN) and if unilateral middle ear ossicle removal led to synaptic loss in the CN. Synaptic endings were identified immunohistochemically, using a monoclonal antibody to synaptophysin. Immunolabeling was quantified densitometrically in the CN 4-161 days after cochlear removal and 161 days after ossicle removal. Fiber degeneration was visualized with the Nauta-Rasmussen silver method. Tissue shrinkage was measured from drawings of CN sections. Compared to the contralateral side, immunolabeling density ipsilaterally was reduced by 4 days in the anterior division of the anteroventral CN (a-AVCN) and by 7 days in the anterior part of the posteroventral CN (a-PVCN). At 7 days, preterminal fiber degeneration was abundant in both areas. These findings were consistent with the loss of cochlear nerve endings and fibers. At later times, immunolabeling density recovered. In the a-AVCN, tissue shrinkage explained approximately half the recovery of staining density; the rest was attributed to synaptogenesis. In the a-PVCN, the entire recovery was attributed to tissue shrinkage. In the polymorphic layer of the dorsal CN, immunostaining density increased transiently at 4 days, while at 7 days preterminal fiber degeneration was abundant. A net loss of synaptic endings was not detected immunohistochemically. The increased immunostaining density may reflect a transient growth of immature processes or presynaptic endings. Ossicle removal produced a deficit in immunolabeling density only in the ipsilateral a-PVCN, without fiber degeneration, suggesting a loss of presynaptic endings or of synaptophysin expression.

Afferent Pathways↗

Modulating calbindin and parvalbumin immunoreactivity in the cochlear nucleus by moderate noise exposure in mice. . A quantitative study on the dorsal and posteroventral cochlear nucleus.

The number of calbindin D-28k and parvalbumin immunoreactive (IR) neurons were characterized on sections from the cochlear nucleus, dorsal cochlear nucleus (DCN) and posteroventral cochlear nucleus (PVCN) using two-dimensional quantification. After noise exposure (6-12 kHz, 2 h, at either 80 dB SPL or 103 dB SPL), the number of calbindin and parvalbumin immunoreactive neurons increased in CBA/CBA mice. Quantitative analysis of calbindin-IR in the PVCN did not show a statistically significant difference between any of the groups, whereas statistically significant differences in calbindin-IR were found in the DCN for the 103 dB and 80 dB group compared to the control group, and 103 dB compared to the 80 dB group, respectively. A statistically significant increase in the number of parvalbumin-IR neurons in the PVCN and the DCN was evident in the 103 dB and 80 dB group compared to the control group, and in the 103 dB compared to the 80 dB group. The data indicate that increasing sound stimulation causes a graded increase in the expression of calcium-binding protein immunoreactivity in the DCN and PVCN neurons and neuropil. This increase of protein expression is due to increased positive immunoreactivity in 'silent' neurons. These findings implicate that these neurons have the possibility to react against trauma and display calbindin or parvalbumin as a rescue event. The ability to map sound-induced calcium-binding protein changes in auditory neurons may be useful in future studies designed for detecting early patterns of neurodegeneration and neuroprotection in the central auditory pathway.

Acoustic Stimulation↗

Collateral projections from single neurons in the dorsal column nucleus to both the cochlear nucleus and the ventrobasal thalamus: a retrograde double-labeling study in the rat.

In the rats which were injected with Fluoro-Gold into the cochlear nucleus and simultaneously with tetramethylrhodamine-dextran amine into the ventrobasal thalamus, a number of neurons in the gracile and the cuneate nuclei of the dorsal column nucleus (DCN) and in the interpolar and caudal subnuclei of the spinal trigeminal nucleus (STN) were double-labeled retrogradely with the both tracers. The results indicated the existence of single DCN/STN-neurons which provided axons to the cochlear nucleus, mainly ipsilaterally, and to the contralateral ventrobasal thalamus.

Animals↗

Glycinergic and GABAergic inputs affect short-term suppression in the cochlear nucleus.

Most cochlear nucleus (CN) neurons exhibit short-term response suppression to a second stimulus in a paired-pulse (click), forward-masking, paradigm. The magnitude of suppression, which appears to be greater than that observed in acoustic nerve, is dependent on the temporal separation and/or relative intensities of the two stimuli. Recent evidence suggests that inhibitory circuitry ending on CN neurons may mediate this response suppression. Using extracellular recordings from single CN neurons, suppression was evaluated using a forward-masking paradigm. Responses to paired acoustic clicks (i.e., a 'masker' followed by an identical 'probe' click) were measured while the time interval between the masker and probe was varied systematically. The role of inhibitory circuitry in forward-masking in the CN was assessed by pharmacologic manipulation of the GABA(A) and glycine(I) (strychnine-sensitive) receptors. Blockade of glycinergic or GABAergic receptors by iontophoretic application of the antagonists, strychnine and bicuculline methiodide, decreased the effects of forward-masking by shortening recovery times of the probe response in 2/3 of the neurons tested. Conversely, agonist application (glycine, and GABA or muscimol) increased the magnitude of suppression and delayed recovery of the probe response relative to control values. These findings suggest that known circuits releasing glycine and/or GABA mediate short-term response suppression in some CN neurons.

Acoustic Stimulation↗

Granule cell activation of complex-spiking neurons in dorsal cochlear nucleus.

Dorsal cochlear nucleus (DCN) principal cells receive, in addition to their well known auditory inputs, various nonauditory inputs via a cerebellar-like granule cell circuit located in the superficial layers of the DCN. Activation of this circuit (granule cell axons make excitatory synapses on the principal cells but also contact inhibitory interneurons that project to the principal cells) produces strong inhibition of the principal cells. Here we investigate the role of cartwheel cells, homologs of cerebellar Purkinje cells, in producing this inhibition. The responses of type IV units (one type of principal cells) and of cartwheel cells were recorded to ortho- and antidromic activation of the granule cells (i. e., by stimulation of their inputs from the somatosensory cuneate and spinal trigeminal nuclei and by direct stimulation of their parallel fiber axons). Cartwheel cells were identified on the basis of recording depth and complex action potential shape. A four-pulse facilitation paradigm (four pulses at 50 msec intervals) was used; this stimulus allows separation of the apparently simple inhibitory somatosensory response of type IV units into a three-component (inhibition-excitation-inhibition) response. As expected, cartwheel cells are excited by granule cell activation; the latencies and four-pulse amplitudes of these responses correspond to the properties of the second, long-latency inhibitory component of type IV responses. The source of the first, short-latency inhibitory response is still unknown. Nevertheless, these results show that cartwheel cells convey inhibitory polysensory information to DCN principal cells.

Animals↗