Octopus Cells Research Articles

Origins and targets of commissural connections between the cochlear nuclei in guinea pigs

Our objective was to identify the origins and targets of axons that project from one cochlear nucleus to the other. First, retrograde tracers were injected into one cochlear nucleus to label commissural cells in the opposite nucleus. In the dorsal cochlear nucleus, a few cells in the deep layers were labeled; they were not further classified according to type. In the ventral cochlear nucleus, all commissural cells that could be classified were multipolar cells. Second, an anterograde tracer was injected into one cochlear nucleus, and the distribution of boutons in the opposite cochlear nucleus was examined. Labeled boutons were present throughout the ventral cochlear nucleus, where they appeared to contact multipolar cells, spherical and globular bushy cells, and octopus cells. In the dorsal cochlear nucleus, labeled boutons were present in the fusiform cell and deep layers and appeared to contact fusiform cells and cells of unknown type. Many labeled terminals were also present in the granule cell regions. Injections into regions associated with high or low frequencies labeled boutons in corresponding regions in the contralateral ventral cochlear nucleus. Third, multiple tracers were used to determine whether cells that project to the inferior colliculus are contacted by commissural axons. Boutons labeled by anterograde transport of one tracer placed in the cochlear nucleus were frequently observed to be apposed to cells that were labeled by retrograde transport of a different tracer placed in the contralateral inferior colliculus. We conclude that commissural projections originate from multipolar cells throughout the ventral cochlear nucleus (and from a small number of cells in the dorsal cochlear nucleus) and make contact with all major cell types of the cochlear nuclei, including at least some of those that project to the inferior colliculus.

Three classes of inhibitory amino acid terminals in the cochlear nucleus of the guinea pig.

Electron microscopic postembedding immunocytochemistry was used to analyze and assess the synaptic distribution of glycine (GLY) and gamma-amino butyric acid (GABA) immunoreactivities in the guinea pig cochlear nucleus (CN). Three classes of endings were identified containing immunolabeling for glycine, GABA, or both glycine and GABA (GLY/GABA). All classes were similar in that the terminals contained pleomorphic vesicles and formed symmetric synapses with their postsynaptic targets. A fourth class, which labeled with neither antibody, contained round vesicles and formed asymmetric synapses. Glycine endings predominated in the ventral CN, while GLY/GABA endings were prevalent in the dorsal CN. GABA endings were the least common and smallest in size. Glycine, GLY/GABA, and GABA endings differed in their proportions and patterns of distribution on the different classes of projection neurons in the CN, including spherical bushy, type I stellate/multipolar, and octopus cells in the ventral CN and fusiform cells in the dorsal CN. The vast majority of anatomically-defined, putative inhibitory endings contain GLY, GABA, or both, suggesting that most of the inhibition in the cochlear nucleus is mediated by these three cytochemically and, probably, functionally distinct classes of endings. The results of this study also suggest that a large proportion of the GABA available for inhibition in the CN coexists in terminals with glycine.

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.

Projections from the cochlear nucleus to the superior paraolivary nucleus in guinea pigs.

Axonal tracing techniques were used to study the projection from the cochlear nucleus to the superior paraolivary nucleus in guinea pigs. Different tracers were used to identify the cell types that give rise to the projections, the morphology of their axons, and the cell types that they contact in the superior paraolivary nucleus. Injections of Fluoro-Gold or peroxidase-labeled-WGA and HRP into the superior paraolivary nucleus labeled multipolar cells and octopus cells bilaterally in the ventral cochlear nucleus, mainly on the contralateral side. Injections of PHAL into the ventral cochlear nucleus labeled two types of axons in the superior paraolivary nucleus. Thin axons branch infrequently and give rise primarily to small, en passant boutons. Thick axons have larger boutons, many of which are terminal boutons that arise from short collaterals. Thin axons appear to originate from multipolar cells, whereas thick axons probably originate from octopus cells. Both types are found bilaterally after an injection into the ventral cochlear nucleus on one side. Individual thick or thin axons may contact multiple cell types in the superior paraolivary nucleus. Individual cells in the superior paraolivary nucleus can receive convergent input from both thick and thin axons. Combined anterograde and retrograde transport of different fluorescent tracers was used to identify the projections of the cells in the superior paraolivary nucleus that receive inputs from the ventral cochlear nucleus. Cells in the superior paraolivary nucleus that projected to the ipsilateral cochlear nucleus or to the ipsilateral inferior colliculus appeared to be contacted by axons that were labeled by anterograde transport from the contralateral ventral cochlear nucleus. Thus the projections to the superior paraolivary nucleus are in a position to affect the activity in both ascending and descending auditory pathways.

Expression of acidic FGF mRNA in rat auditory brainstem during postnatal maturation

In situ hybridization was used to investigate the mRNA distribution of acidic and basic fibroblast growth factor (aFGF and bFGF) in the auditory brainstem of neonatal and adult rats. bFGF mRNA was not detected at any age. In adult rats, aFGF mRNA was strongly expressed in the principal neurons of the anteroventral and posteroventral cochlear nuclei, but not in the octopus cells. In the dorsal cochlear nucleus, aFGF mRNA was seen only in scattered smaller vertical cells. aFGF was strongly expressed in the nucleus of the trapezoid body and in all periolivary cell groups, but not in the medial and lateral olivary nuclei. No expression was observed in the lemniscal nuclei or in the central nucleus of the inferior colliculus, but large neurons in the external zone of the colliculus were labeled. Developmentally, low levels of aFGF expression appeared in the cochlear nuclei and olivary nuclei between P0 and P6. This expression increased rapidly during the onset of hearing, between P10 and P14, and reached adult level by P14–P17. Labeling in collicular neurons appeared slightly later. The results suggest that the appearance of strong aFGF mRNA expression is related to the onset of function.

Recordings from slices indicate that octopus cells of the cochlear nucleus detect coincident firing of auditory nerve fibers with temporal precision

Acoustic information in auditory nerve discharges is integrated in the cochlear nuclei, and ascends through several parallel pathways to higher centers. Octopus cells of the posteroventral cochlear nucleus form a pathway known to carry information in the timing of action potentials. Octopus cells have dendrites oriented to receive converging input from many auditory nerve fibers. In all 34 intracellular recordings from anatomically identified octopus cells in slices, shocks to the auditory nerve evoked brief, consistent, graded EPSPs. EPSPs were about 1 msec in duration. At all but the lowest shock strengths, the delays between shocks and the peaks of resultant EPSPs had SDs of 0.02 msec. Polysynaptic excitation, perhaps arising from the axon collaterals of octopus cells, was observed. No detectable glycinergic or GABAergic inhibition was evoked with shocks. The input resistances were low, around 10 M omega, voltage changes were rapid, with time constants of about 1 msec, and action potentials were small. The low input resistance resulted in part from a Cs(+)-sensitive conductance. In the presence of 10 or 15 mM extracellular Cs+ the time constants increased 20-fold in the hyperpolarizing voltage range. As several subthreshold inputs were required to produce suprathreshold responses, octopus cells detect the coincident firing of auditory nerve fibers. Under physiological conditions the low input resistance and resulting short time constant limit the time over which temporal summation of excitation from auditory nerve fibers can occur and thus provide temporal precision to electrical signaling.

Morphology of the cochlear nucleus in CBA/J mice with chronic, severe sensorineural cochlear pathology induced during adulthood

The effects of chronic cochlear impairment on morphological features of the adult cochlear nucleus (CN) were assessed in CBA/J mice in which severe sensorineural damage had been induced by exposure to intense noise. Sections from various CN subdivisions, stained for Nissl substance and fibers, were quantitatively evaluated in four groups of noise-exposed mice that differed with regard to the age at noise exposure (2, 6, or 11 months), age at the time the CN was evaluated (6, 11, or 24 months), and the duration (chronicity) of sensorineural impairment (4, 5, 13, or 18 months). Like-aged, non-exposed CBA mice were used as controls, so the effects of peripheral damage and aging could be compared. Cochlear damage produced significant changes in CN subdivisions thought to receive the heaviest input from cochlear afferents (anteroventral CN, octopus cell area, dorsal CN layer III). These changes included a reduction of neuropil volume, reductions in neuron size, and increases in neuronal packing density that were complementary to reduced volume in these subdivisions. Effects on neuron number were minimal in all subdivisions. Central changes in noise-exposed mice were absent or diminished in DCN layers I and II, which receive relatively less input from primary fibers. The age at onset and chronicity of damage had little to do with the severity of central effects of cochlear damage. The effects of cochlear damage were not additive with age-related changes seen in the old controls.

Central projections of auditory nerve fibers of differing spontaneous rate, II: Posteroventral and dorsal cochlear nuclei

Response properties of auditory nerve fibers (ANFs), including threshold sensitivity, vary systematically with spontaneous discharge rate (SR) (Liberman, M.C.: J. Acoust. Soc Amer. 63:442-455, 1978). Thus, an understanding of the mechanisms underlying signal transformation in the cochlear nucleus (CN) must include a description of any SR-based difference in ANF projections. This study is the second of a pair describing the CN projections of intracellularly labeled ANFs of known SR, the first of which summarized projection to the anteroventral CN (Liberman, M.C.: J. Comp. Neurol. 313:240-258, 1991). For each swelling from each labeled fiber, the position (within CN subdivisions), the size, and the type of cell contacted (if determinable) was noted: roughly one in four labeled swellings appeared in intimate contact with the soma or proximal dendrites of a CN cell. In all such cases, cell size and swelling size were measured. As reported for auteroventral cochlear nucleus, the ANF innervation of the small-cell regions of posteroventral CN (PVCN) was almost exclusively by low- and medium-SR fibers. Other significant SR-based trends in ANF projections included 1) a tendency for high-SR fibers to contact larger cells in PVCN, 2) a meager projection of low- and medium-SR fibers to octopus cells, and 3) a tendency in the dorsal CN (DCN) for low-SR terminals to end closer to the fusiform cell layer than high-SR terminals. There were no significant SR-based difference in ANF swelling sizes in any subdivision. A consideration of the average cell sizes, ANF swelling sizes and estimated numbers of ANFs of different CF and SR converging on each CN cell help explain some of the differences in response transformation associated with different cell types in the CN.

Tuberculoventral neurons project to the multipolar cell area but not to the octopus cell area of the posteroventral cochlear nucleus

Tuberculoventral neurons in the deep layer of the dorsal cochlear nucleus (DCN) provide frequency-specific inhibition to neurons in the anteroventral cochlear nucleus (AVCN) of the mouse (Wickesberg and Oertel, '88, '90). The present experiments examine the projection from the deep DCN to the posteroventral cochlear nucleus (PVCN). Horseradish peroxidase (HRP) injections into the PVCN reveal that the multipolar cell area, but not the octopus cell area, is innervated by neurons in the deep layer of the DCN. Injections into the multipolar cell area, in the rostral and ventral PVCN, labeled neurons across the entire rostrocaudal extent of the deep DCN. The labeled tuberculoventral neurons generally lay within the band of labeled auditory nerve terminals in the DCN. Injections of HRP into the octopus cell area, in the dorsal caudal PVCN, labeled almost no cells within the band of auditory nerve fiber terminals that were labeled by the same injection. The inhibition from tuberculoventral neurons onto ventral cochlear nucleus (VCN) neurons is likely to be mediated by glycine (Wickesberg and Oertel, '90). Slices of the cochlear nuclear complex were immunolabeled by an antibody against glycine conjugated with glutaraldehyde to bovine serum albumin (Wenthold et al., '87). Glycine-like immunoreactivity was found throughout the DCN, the AVCN and the multipolar cell area, but there was little labeling in the octopus cell area. This finding provides independent evidence that tuberculoventral neurons do not innervate the octopus cell area and indicates that the octopus cell area is anatomically and functionally distinct.

Calbindin‐like immunoreactivity in the central auditory system of the mustached bat, Pteronotus parnellii

With the aid of a polyclonal antibody specific for Calbindin D-28k, we studied the distribution of this calcium-binding protein in the central auditory system of the mustached bat, Pteronotus parnelli. Components of the cochlear nucleus (CN) that were calbindin-positive (cabp(+] included the root of the auditory nerve, multipolar and globular bushy cells in the anteroventral CN, multipolar and octopus cells in the posteroventral CN, and small and medium-size cells in the dorsal CN. Not stained were spherical bushy cells of the anteroventral CN and pyramidal/fusiform cells in the dorsal CN. In the superior olivary complex, labeled cells were found in the lateral and medial nuclei of the trapezoid body, the ventral and ventromedial periolivary nuclei, and the anterolateral periolivary nucleus. No cellular labeling was seen in the lateral superior olive. In the medial superior olive, only marginal cells were cabp(+). Labeled fibers could be seen surrounding the gosts of unlabeled cells in both the latter nuclei. Most cells in the intermediate nucleus and the columnar division of the ventral nucleus of the lateral lemniscus were cabp(+). However, the dorsal nucleus was cabp(-). A group of cabp(+) cells was also seen in the paralemniscal zone. The inferior colliculus had a relatively low density of cabp(+) cells. Labeled cells were more common in the caudal half of the central nucleus, and in the external nucleus and dorsal cortex. In the auditory thalamus, nearly every cell in the medial geniculate body was cabp(+), but those in the suprageniculate nucleus and in the posterior group did not stain. Small cells in the intermediate layer and giant cells in the deep layers of the superior colliculus were densely cabp(+). In the pons, cabp(+) cells and neuropil could be seen in the medial and lateral pontine nuclei (pontine gray). In conclusion, calbindin-like immunoreactivity was found in most of the brainstem auditory system, as well as in regions associated with acoustic orientation or control of vocalization. However, except for a minority of cells of the medial superior olive, it is conspicuously absent from the nuclei receiving binaural input below the level of the inferior colliculus.

Saccular afferents to second-order cochlear neurons

Previous studies have described a primary afferent vestibular projection to second-order cochlear neurons originating from the saccular maculae. As could be shown in the guinea pig by means of anterograde transport of horseradish peroxidase and immuno-cytochemistry, these saccular afferents terminated at cells immunoreactive to glutamate and aspartate but not to gamma-aminobutyric acid (GABA). These were intermingled among the fibers of the acoustic striae and situated between the dorsal cochlear nucleus and the octopus cell area. Within the acoustic striae, these saccular afferents appear to have a close relationship to GABA-immunoreactive structures, such as boutons en passant and terminal boutons. This striking relationship, along with cytoarchitectural criteria and the saccular input to these second-order cochlear neurons, gave reason to discern them as a separate cell group.

Morphology of the octopus cell area of the cochlear nucleus in young and aging C57BL/6J and CBA/J mice

The influence of aging and age-related cochlear impairment on the ventral cochlear nucleus was evaluated by measuring morphological properties of the octopus cell area (OCA) in five age groups of inbred C57BL/6J and CBA/J mice (young adult to very old). The former strain demonstrates progressive cochlear sensorineural pathology and hearing loss during middle age; the latter has only modest sensorineural pathology late in life. Histological sections of the OCA were evaluated with serial sections and several strains for neurons, glia, and fibers, and Golgi impregnations were also used. Aging was associated with a decrease in volume of the OCA, a loss of neurons, slight decrease in neuron size, increased packing density of glial cells, and changes in dendrites ranging from minor to total loss of primary branches. The greatest changes occurred in extreme old age, beyond the median lifespan. Age-related changes were not exacerbated by sensorineural pathology in aging C57BL/6J mice. Individual octopus cells varied greatly in the extent of age-related abnormality.

Anatomy of the cochlear nuclear complex of guinea pig

The cyto- and fibre-architecture of the cochlear nuclear complex of the guinea-pig has been studied in serial sections using Nissl, Golgi and combined cell-myelin staining of normal material, and a silver degeneration method after cochlear ablation. The nuclear subdivisions and major cell types can be recognised on the basis of those found in the cat, but there are some differences between the two species in the precise distribution and morphology of the neurons. The rostrodorsal part of the anteroventral cochlear nucleus (AVCN) contains predominantly spherical bushy cells, but these cannot be readily divided into large and small types as in the cat. Globular bushy cells are seen in the caudal region of the AVCN, but the majority occur in the posteroventral cochlear nucleus (PVCN), in an area extending from the nerve root right up to the boundary of the dorsal cochlear nucleus (DCN). The octopus cells constitute a distinct region in the most dorsomedial part of the PVCN underneath the DCN. Giant cells are seen scattered around the nerve root region. Multipolar and small cells are seen throughout the non-granular regions of the ventral cochlear nucleus (VCN) except for the octopus cell area, but occur mainly in the more rostral regions of the PVCN. Small cells occur in greatest abundance in the thin cap area at the dorsal edge of the VCN below a superficial granule cell layer. The latter covers the dorsolateral surface of the VCN, and a lamina of granule cells partially separates the PVCN from the DCN. The DCN can be divided into four layers. The outermost molecular layer (layer 1) is separated from the deeper regions by a prominent layer of granule cells (layer 2) which also contains the pyramidal cells. Molecular layer stellate cells are seen in layer 1 and a staggered row of cartwheel neurons is found at the boundary between layers 1 and 2. Layer 3 contains the basal dendrites of the pyramidal cells and some small (vertical) cells, and is innervated by the descending branches of the cochlear nerve. The deepest layer 4, which contains multipolar cells and giant cells, does not appear to receive this direct cochlear input.

Morphology and physiology of cells in slice preparations of the posteroventral cochlear nucleus of mice

In an effort to understand what integrative tasks are performed in the cochlear nuclei, the present study was undertaken to describe neuronal circuits in the posteroventral cochlear nucleus (PVCN) anatomically and physiologically. The cochlear nuclear complex receives auditory information from the cochlea through the auditory nerve. Within the cochlear nuclei, signals travel along several parallel and interconnected pathways. From the cochlear nuclei, transformed versions of the signals are passed to higher auditory centers in the brainstem. We have recorded electrophysiological responses from cells that were subsequently visualized with horseradish peroxidase (HRP). Responses to shocks to the auditory nerve root and to intracellularly injected current pulses were recorded and correlated with morphology. Two types of stellate cells and octopus cells were distinguished. T stellate cells project out of the cochlear nuclei through the Trapezoid body; D stellate cells do not. The axons of D stellate cells extend Dorsalward to the dorsal cochlear nucleus (DCN) but have not been traced out of the nucleus. Both T and D stellate cells have terminal collaterals in the multipolar cell region of the PVCN and in the DCN. The endings of one T stellate cell formed a narrow band rostrocaudally in the fusiform cell layer of the DCN that resembled an isofrequency band. The endings of one D stellate cell lay closely apposed to multipolar cells in the deep layer of the DCN. The dendrites of T stellate cells are often aligned along the path of auditory nerve fibers and end in tufts, whereas those of D stellate cells extend radially in the plane of the lateral surface of the PVCN toward granule cell areas and branch sparingly. Octopus cells have dendrites oriented perpendicularly to the path of auditory nerve fibers. Their axons were cut medially in the slices; none had collateral branches. Both T and D stellate cells were monosynaptically excited to threshold by shocks to the nerve root, indicating that they could participate in local circuits that we measure physiologically. T stellate cells have action potentials that peak at about 0 mV and are followed by single undershoots. The D stellate cell that was best impaled fired overshooting action potentials that were followed by double undershoots. Octopus cells were monosynaptically excited to threshold by shocks to the auditory nerve.

The form and distribution of GABAergic synapses on the principal cell types of the ventral cochlear nucleus of the cat

The distribution of GABAergic endings was examined histochemically in the ventral cochlear nucleus (VCN) of the cat using an antibody to glutamate decarboxylase (GAD), the synthetic enzyme for GABA. Immunoreactive (GAD +) endings appeared in all subdivisions of the cat VCN. Each of the principal cell types had a characteristic labeling pattern, based on the size, concentration, and distribution of GAD + endings on its soma. Spherical bushy cell somata were typically contacted by many small (<1.5 μm in diameter) and medium-sized (1.5–2 μm in diameter) endings, many of which aggregated into tight clusters. Globular bushy cells had a similar pattern, but the clusters of GAD + endings were less tightly packed. Reactive endings on stellate cells were more evenly distributed. GAD + endings on octopus cells were larger (up to 2.5 μm in diameter) than those on the bushy cells and tended to aggregate into small clusters or rows on the somata and dendrites. Reactive endings contained small pleomorphic vesicles and formed symmetrical synaptic contacts on each of the cell types examined. The patterns formed by GAD + endings on each type of neuron resemble those of certain types of non-cochlear axons previously described with the Golgi methods as projecting from the dorsal cochlear nucleus and the trapezoid body.

Divergent projections of physiologically characterized rat ventral cochlear nucleus neurons as shown by intra-axonal injection of horseradish peroxidase.

An attempt was made to correlate electrophysiological and morphological characteristics of rat ventral cochlear nucleus neurons. Their axonal course and their soma morphology were investigated using the intra-axonal horseradish peroxidase method. Prior to labeling, neurons were characterized by recording their response patterns to acoustic stimulation with pure tones. Three types of cells were found: Category I (37 neurons) exhibited "primarylike" responses and a spontaneous firing rate below 10 spikes/s. Category II (21 neurons) showed "on" responses and little spontaneous activity. Category III (9 neurons) had "primarylike" responses like neurons in category I. However, the spontaneous activity rate of these neurons was significantly higher (mean: 95 spikes/s). Among the response categories, the morphological characteristics differed in some prominent aspects. Within each category, however, the morphological properties were rather similar. All neurons in category I were globular/bushy cells located in the area of the entrance of the cochlear nerve. The axon of each cell coursed along the ventral acoustic stria and consistently innervated the lateral superior olive ipsilaterally, and the nucleus of the trapezoid body and the nucleus of the lateral lemniscus contralaterally. Some neurons also projected to periolivary nuclei ipsilaterally and contralaterally. Neurons in category II were located in the posteroventral cochlear nucleus and were presumably multipolar/stellate cells. Their axons coursed via the intermediate acoustic stria and innervated mainly contralateral periolivary regions as well as the contralateral nucleus of the lateral lemniscus. Ipsilaterally, the lateral superior olive and the superior periolivary nucleus were innervated by some of the category II neurons. Somata types of neurons in category III could not be identified morphologically, but somata were located in caudal parts of the posteroventral cochlear nucleus that correspond to the octopus cell area. Their axons coursed via the intermediate acoustic stria and innervated periolivary regions and the contralateral nucleus of the lateral lemniscus. Thus, their axonal distribution differed only slightly from neurons in category II. These data confirm and extend previous findings regarding the efferent connections of ventral cochlear neurons. They emphasize the complexity of the axonal projection patterns of single cochlear nucleus cells. Since two types of response patterns and three types of axonal projection patterns have been observed, there remains an ambiguous relation between response pattern and axonal projection site.(ABSTRACT TRUNCATED AT 250 WORDS)

Glycine receptor immunoreactivity in the ventral cochlear nucleus of the guinea pig.

Glycine appears to be a major inhibitory neurotransmitter in the cochlear nucleus. In order to determine more precisely the distribution of glycinergic synapses, we have studied the immunocytochemical distribution of the glycine postsynaptic receptor. Two monoclonal antibodies were used, Gly Rec Ab 2, which recognizes the 48kD polypeptide and Gly Rec Ab 7, which primarily recognizes the 93kD subunit of the glycine receptor complex. At the light microscopic level, glycine receptor immunoreactivity was found throughout the ventral cochlear nucleus with a punctuate distribution often found outlining large cell bodies. Indistinguishable patterns of staining were obtained with the two antibodies. Ultrastructural localization was done with Gly Rec Ab 7 because immunoreactivity remained after fixation with glutaraldehyde containing solutions. At the ultrastructural level, immunoreactivity was concentrated at postsynaptic sites on dendrites and cell bodies. In the anteroventral cochlear nucleus, neurons identified as spherical cells contained numerous inmunoreactive synapses on their cell bodies, whereas most immunoreactive synapses on stellate cells were on their proximal dendrites. In the posteroventral cochlear nucleus, neurons identified as octopus cells were immunoreactive on their cell bodies and proximal dendrites. In the granule cell layer, immunoreactivity was found only in the neuropile. Throughout the ventral cochlear nucleus, glycine receptor immunoreactivity was found postsynaptic to terminals containing flattened synaptic vesicles as well as those containing oval/pleomorphic synaptic vesicles.

Patterns of glutamate decarboxylase immunostaining in the feline cochlear nuclear complex studied with silver enhancement and electron microscopy

The cochlear nuclear complex of the cat was immunostained with an antiserum to glutamate decarboxylase (GAD), the biosynthetic enzyme for the inhibitory neurotransmitter GABA, and studied with different procedures, including silver intensification, topical colchicine injections, semithin sections, and immunoelectron microscopy. Immunostaining was found in all portions of the nucleus. Relatively few immunostained cell bodies were observed: most of these were in the dorsal cochlear nucleus and included stellate cells, cartwheel cells, Golgi cells, and unidentified cells in the deep layers. An accumulation of immunoreactive cells was also found within the small cell cap and along the medial border of the ventral cochlear nucleus. Immunostained cells were sparse in magnocellular portions of the ventral nucleus. Most staining within the nucleus was of nerve terminals. These included small boutons that were prominent in the neuropil of the dorsal cochlear nucleus, the granule cell domain, in a region beneath the superficial granule cell layer within the small cell cap region, and along the medial border of the ventral nucleus. Octopus cells showed small, GAD-positive terminals distributed at moderate density on both cell bodies and dendrites. Larger, more distinctive terminals were identified on the large cells in the ventral nucleus, in particular on spherical cells and globular cells. There was a striking positive correlation of the size, location, and complexity of GAD-positive terminals with the size, location, and complexity of primary fiber endings on the same cells. This correlation did not hold in the dorsal nucleus, where pyramidal cells receive many large GAD-positive somatic terminals despite the paucity of primary endings on their cell bodies. The GAD-positive terminals contained pleomorphic synaptic vesicles and formed symmetric synaptic junctions that occupied a substantial portion of the appositional surface to cell bodies, dendrites, axon hillocks, and the beginning portion of the initial axon segments. Thus, the cells provided with large terminals can be subjected to considerable inhibition that may be activated indirectly through primary fibers and interneurons or by descending inputs from the auditory brainstem.

The central projections of intracellularly labeled auditory nerve fibers in cats.

The central projections of physiologically characterized auditory nerve fibers were studied in the cochlear nuclei of adult cats after intracellular staining with horseradish peroxidase (HRP). This technique consistently labels only the type I spiral ganglion neurons which contact inner hair cells in the cochlea (Liberman and Oliver, '84). The central axon of each type I neuron bifurcates in the cochlear nucleus to form an ascending branch and a descending branch. The characteristic frequency (CF) of a fiber corresponds to the dorsoventral position of these major branches and their collateral ramifications within the nucleus. Fibers of low CFs are distributed ventrally, and fibers of increasing CF are distributed progressively more dorsally. In some cases, the collateral branches deviate from this tonotopic arrangement, particularly in (1) the octopus cell region of the posteroventral cochlear nucleus, (2) the zone of bifurcations of the auditory nerve fibers, and (3) the anterior, dorsal, and lateral margins of the ventral cochlear nucleus. Spontaneous discharge rate (SR) is related to the complexity of the axon arbor, especially along the ascending branch. Fibers of low and medium SR exhibit more axonal branch points and longer collateral lengths than do those with high SR. Six of 37 labeled fibers fail to innervate the dorsal cochlear nucleus, a feature apparently unrelated to CF or SR.

Effect of neonatal conductive hearing loss on neuronal size in the ventral cochlear nucleus of guinea pigs

The left ear of newborn guinea pigs was closed by removing the skin and cartilage of the external auditory meatus and pulling the surrounding tissue over the opening with silk sutures. At 60 days of age the animals were anesthetized with chloral hydrate and auditory brainstem responses (ABR) were measured before and after destroying the intact right ear. After ABR testing the animals were killed and their cochleas were examined histologically. Right and left brainstems were embedded in paraffin, serially sectioned and stained with cresyl violet. The cross‐sectional area of four different cell types was measured, using a total of 210 cells of each type from each side. For all four groups; large spherical cells, small spherical cells, pale globular cells, and octopus cells, soma size was significantly smaller (P < 0.01) on the left side. This result for guinea pigs, a precocial animal, agrees with earlier findings for an altrical animal the mouse [D. B. Webster, Exp. Neurol. 79, 130–140 (1983)]. [Supported by NIH training grant NSD‐07058 and the Louisiana Lions Eye Foundation.] Present address: Dept. of Biological Sciences, Loyola University, New Orleans, LA 70118.