Synaptic connections of the auditory nerve in cats: relationship between endbulbs of held and spherical bushy cells

This report focuses on a class of large synaptic endings, the endbulbs of Held. These endings are located in the anteroventral cochlear nucleus and arise from the axons of type I spiral ganglion neurons. Axons were stained with horseradish peroxidase (HRP) using intracellular injections of single fibers or extracellular injections into the auditory nerve. Individual endbulbs or pairs of endbulbs that converged onto the same spherical bushy cell were examined with the aid of a light microscope and subjected to morphometric analyses. Endbulbs of fibers having low spontaneous discharge rates (SR, less than or equal to 18 spikes/sec) have a more complex shape than those of high SR fibers (greater than 18 s/s), a feature represented by systematic differences in endbulb silhouette perimeter without differences in endbulb silhouette area. Consequently, the ratio, silhouette area divided by silhouette perimeter, yields a "form factor" separating endbulbs of high SR from those of low SR. High SR fibers had ratios greater than 0.52 (mean = 0.63 +/- 0.09), whereas low SR fibers had ratios less than 0.52 (mean = 0.45 +/- 0.06). Pairs of endbulbs with unknown physiological properties had similar form factor values, despite the wide range of values observed in the endbulb population. These data imply that endbulbs converging upon the cell body of a spherical bushy cell arise from fibers of the same SR group. Electron microscopic examination was conducted on the endbulb of one physiologically characterized and intracellularly stained auditory nerve fiber (CF = 1.4 kHz; SR = 55 s/s) and its unstained endbulb mate with the aid of serial ultrathin sections. In addition to the well-known axosomatic synapses, these endbulbs formed axodendritic synapses: 11.7% for the HRP-labeled endbulb and 13.3% for the unlabeled endbulb. The axodendritic synapses appear to occur on dendrites of nearby spherical bushy cells and may represent a mechanism whereby single endbulbs can disperse activity to multiple neurons in the cochlear nucleus. We propose that axosomatic synapses preserve fiber SR groupings, whereas axodendritic synapses may not.     

Selective labeling of spiral ganglion and granule cells with D-aspartate in the auditory system of cat and guinea pig

The present study sought to locate putative glutamatergic or aspartatergic pathways in the auditory system of cats and guinea pigs. We injected 0.06 to 3 mM D-[3H] aspartate (D-Asp) in the cochlear nucleus before preparation for light microscopic autoradiography. At short survival times (15 and 40 min) there was heavy labeling of astrocytic somata. Labeling patterns typical of cochlear nerve endings decorated neurons in the cochlear nucleus, e.g., cell bodies and dendritic trunks of octopus cells. Labeling patterns consistent with retrograde axonal transport by the parallel fibers of granule cells appeared in the molecular layer of the dorsal cochlear nucleus and in the external granular layer. Retrograde labeling of the cochlear nerve root fibers also occurred. Consistent with these results are companion biochemical findings on the rapidly dissected cochlear nuclei of guinea pigs. The dorsal, anteroventral, and posteroventral cochlear nuclei, each, evinced uptake of D-Asp. Subsequently, electrical stimulation of each nucleus released a portion of the accumulated amino acid. Most of this release probably came from synaptic endings. Another group of experiments compared autoradiographic localization of 0.06 to 3 mM D-Asp to that of horseradish peroxidase (HRP) 6 hr to 2 d after injections in the cochlear nucleus. Astroglial cell bodies were no longer labeled by D-Asp, but spiral ganglion cell bodies in the cochlea and granule cell bodies in the cochlear nucleus were. Perikarya of the periolivary and ventral cochlear nuclei projecting to the dorsal cochlear nucleus were labeled by HRP and not by D-Asp. Thus, comparisons with the HRP findings indicate that D-Asp labeling resulted from a selective retrograde transport. There was no evidence for a selective anterograde axonal transport. The present observations support the hypothesis that cochlear nerve fibers and granule cells may use L-glutamate and/or L-aspartate as a transmitter in the cochlear nucleus.     

Endbulbs of held and spherical bushy cells in cats: morphological correlates with physiological properties

Single auditory nerve fibers of type I spiral ganglion cells in cats were electrophysiologically characterized by recording with micropipettes inserted into the axon and then labeled by intracellular injections of horseradish peroxidase (HRP) through the same pipettes. This method for staining and studying single neurons allowed us to describe structure-function relationships for labeled endbulbs of Held and the somata of their postsynaptic spherical bushy cells. The silhouette areas of terminal endbulbs and the corresponding somata of spherical bushy cells were determined by planimetry from drawings made with a light microscope and drawing tube. On the presynaptic side, endbulb area is related to fiber characteristic frequency (CF, the frequency to which a fiber is most sensitive) such that the largest endbulbs arise from fibers having CFs between 1 and 4 kHz; smaller endbulbs can arise from fibers of any CF. Endbulb area is not correlated with fiber spontaneous discharge rate (SR). Dividing the endbulb's silhouette area by its silhouette perimeter, however, yields a "form factor" that is a reliable indicator of fiber SR: Endbulbs from fibers of low-medium SR (less than or equal to 18 spikes/second) have form factor values less than 0.52, whereas endbulbs of high SR fibers (greater than 18 spikes/second) have values greater than 0.52. This form factor should therefore be predictive of SR groupings in auditory fibers for which physiological data are not available. On the postsynaptic side, the somata of spherical bushy cells receiving endbulbs from low-medium SR fibers are on average smaller than those receiving endbulbs from high SR fibers. In contrast, the nuclei of the spherical bushy cells are the same size regardless of presynaptic fiber SR. Some of the effects of low-medium SR fibers on their postsynaptic targets, when compared to those of high SR fibers, appear to be mimicked by effects of experimentally induced deprivation.     

Tonotopic projection from the dorsal to the anteroventral cochlear nucleus of mice

To understand how auditory information is processed in the cochlear nuclei, it is crucial to know what circuitry exists and how it functions. In slice preparations, horseradish peroxidase (HRP) injections into the anteroventral cochlear nucleus (AVCN) reveal two circuits: a connection between the dorsal cochlear nucleus (DCN) and AVCN and a local circuit confined to the AVCN. Extracellular injection in the AVCN labels a band of cells in the DCN. The labeled cells in the DCN lie within a band of auditory nerve fiber terminals that are labeled by the same injection, showing that the connection from the DCN to the AVCN is frequency specific. The injections into the AVCN also labeled a cluster of neurons in the AVCN dorsal to the injection site. These cells may be interneurons that relay information from areas encoding higher frequencies to areas encoding lower frequencies within the AVCN. In the parasagittal plane, the AVCN is organized along two orthogonal axes that are indicated with HRP labeling of fibers and cell bodies. The tonotopic axis runs approximately dorsoventrally; the isofrequency axis runs approximately rostrocaudally. The axons of labeled DCN neurons and the cluster lie along the tonotopic axis, whereas the labeled auditory nerve fibers define the isofrequency axis. Where they cross is where HRP is taken up by the fibers. The area of uptake is small and lies in the middle of the darkly stained injection site.     

Selective retrograde labeling of lateral olivocochlear neurons in the brainstem based on preferential uptake of 3H-D-aspartic acid in the cochlea

We have previously shown that perfusion of the gerbil cochlea with probe concentrations of 3H-D-aspartic acid (D-ASP) results in immediate, selective labeling of 50-60% of the efferent terminals under the inner hair cells, presumably by high-affinity uptake. The present study was undertaken to determine the origin of these endings. Twenty-four hours after cochlear perfusion with D-ASP, labeled neurons were observed in the ipsilateral, and to a much lesser extent in the contralateral, lateral superior olivary nucleus (LSO). The cells were small, primarily fusiform, and showed fewer synaptic contacts than other LSO cells. Combined transport of D-ASP and horseradish peroxidase indicated that all olivocochlear neurons within the LSO that projected to the injected cochlea were labeled by D-ASP. Labeled fibers coursed dorsally from the LSO, joined contralateral fibers that had passed under the floor of the fourth ventricle, and entered the VIIIth nerve root at its ventromedial edge. Adjacent to the ventral cochlear nucleus (VCN), densely labeled collateral fibers crossed the nerve root to enter the VCN. Labeled fibers and terminals were prominent in the central VCN. Neither retrograde transport of D-ASP by medial olivocochlear and vestibular efferents nor anterograde transport by VIIIth nerve afferents was observed. The D-ASP-labeled cells and fibers are clearly lateral olivocochlear efferents. Retrograde transport of D-ASP thus allows the cells, axons, and collaterals of the lateral olivocochlear system to be studied, morphologically, in isolation from other cells that project to the cochlea. Since the olivocochlear neurons are almost certainly cholinergic, retrograde amino acid transport does not necessarily identify the primary neurotransmitter of a neuron. Rather, it indicates the presence of selective uptake by the processes of that neuron at the site of amino acid injection. Retrograde labeling appears to be markedly enhanced by the use of metabolically inert compounds such as d-isomer amino acids.     

Unmyelinated axons of the auditory nerve in cats.

This paper describes some central terminations of type II spiral ganglion neurons as labeled by extracellular injections of horseradish peroxidase (HRP) into the auditory nerve of cats. After histological processing with diaminobenzidine, both thick (2-4 microns) and thin (0.5 microns) fibers of the auditory nerve were stained. Whenever traced, thick fibers always originated from type I spiral ganglion neurons and thin fibers always from type II ganglion neurons. Because the labeling of type II axons faded as fibers projected into the cochlear nucleus, this report is limited to regions of the ventral cochlear nucleus near the auditory nerve root. The central axons of type II neurons are unmyelinated, have simple yet variable branching patterns in the cochlear nucleus, and form both en passant and terminal swellings. Under the light microscope, most swellings are located in the neuropil but they are also found in the vicinity of cell bodies, nodes of Ranvier of type I axons, and blood vessels. Eighteen en passant swellings in the neuropil were located by light microscopy and resectioned for electron microscopy; two of these swellings exhibited ultrastructural features characteristic of chemical synapses. The data indicate that inputs from outer hair cells might be able to influence auditory processing in the cochlear nucleus through type II primary neurons.     

Single unit recordings in the auditory nerve of congenitally deaf white cats: Morphological correlates in the cochlea and cochlear nucleus

It is well known that experimentally induced cochlear damage produces structural, physiological, and biochemical alterations in neurons of the cochlear nucleus. In contrast, much less is known with respect to the naturally occurring cochlear pathology presented by congenital deafness. The present study attempts to relate organ of Corti structure and auditory nerve activity to the morphology of primary synaptic endings in the cochlear nucleus of congenitally deaf white cats. Our observations reveal that the amount of sound-evoked spike activity in auditory nerve fibers influences terminal morphology and synaptic structure in the anteroventral cochlear nucleus. Some white cats had no hearing. They exhibited severely reduced spontaneous activity and no sound-evoked activity in auditory nerve fibers. They had no recognizable organ of Corti, presented >90% loss of spiral ganglion cells, and displayed marked structural abnormalities of endbulbs of Held and their synapses. Other white cats had partial hearing and possessed auditory nerve fibers with a wide range of spontaneous activity but elevated sound-evoked thresholds (60–70 dB SPL). They also exhibited obvious abnormalities in the tectorial membrane, supporting cells, and Reissner's membrane throughout the cochlear duct and had complete inner and outer hair cell loss in the base. The spatial distribution of spiral ganglion cell loss correlated with the pattern of hair cell loss. Primary neurons of hearing-impaired cats displayed structural abnormalities of their endbulbs and synapses in the cochlear nucleus which were intermediate in form compared to normal and totally deaf cats. Changes in endbulb structure appear to correspond to relative levels of deafness. These data suggest that endbulb structure is significantly influenced by sound-evoked auditory nerve activity. J. Comp. Neurol. 397:532–548, 1998. © 1998 Wiley-Liss, Inc.     

Morphology of labeled afferent fibers in the guinea pig cochlea

Cochlear afferent and efferent fibers in the guinea pig were labeled by focal extracellular injections of horseradish peroxidase into the spiral ganglion of the basal turn. The morphology and pattern of termination of these fibers were studied by light microscopy. Fibers labeled by injections into the peripheral side of the ganglion could be grouped on the basis of their courses and terminations in the cochlea into two classes of afferent fibers, two classes of efferent (olivocochlear) fibers, and other presumably autonomic fibers. This paper describes the characteristics of labeled afferent fibers and their parent ganglion cells. Peripheral afferent fibers were grouped into two major classes: thick (mean diameter 1.7 micron) radial fibers projecting in a primarily radial fashion from the spiral ganglion and terminating on single inner hair cells and thin (mean diameter 0.5 micron) outer spiral fibers that spiral basalward in the organ of Corti to terminate on outer hair cells, usually in one row. For outer spiral fibers, the number of outer hair cells contacted and the length of the terminal region depend on the row of outer hair cells contacted, with third-row fibers forming, on the average, the most extensive region of termination. Within the spiral ganglion, two types of ganglion cells could be distinguished: type-I ganglion cells of large size (mean soma area = 216 microns 2) with a ratio of central process diameter to peripheral process diameter greater than one and type-II ganglion cells of smaller size (mean soma area = 100 microns 2) and a central to peripheral process ratio near one. In three cochleae in which injections were made central to the ganglion, 11 type-I ganglion cells have been traced to radial fibers contacting inner hair cells and eight type-II ganglion cells have been traced to outer spiral fibers contacting outer hair cells. Thus the afferent innervation of the guinea pig cochlea is similar to the pattern described in other mammals, in which there is separate innervation of the inner and outer hair cells by the two types of ganglion cells. The central axons of both types of ganglion cells were traced individually through serial sections of a block of tissue containing the cochlea, the auditory nerve, and the cochlear nucleus. They followed similar courses in the auditory nerve, and the axons followed into the cochlear nucleus bifurcated in similar regions of the interstitial portion.(ABSTRACT TRUNCATED AT 400 WORDS)     

Distribution of primary vestibular fibers in the brainstem and cerebellum of the monkey

Attempts were made to determine the central projections of ganglion cells innervating individual semicircular ducts in the monkey by implanting or injecting tritiated amino acids (leucine and/or proline), or horseradish peroxidase (HRP), selectively into a single ampulla. Central transport via the vestibular ganglion in animals receiving isotope implants or injections fell into three categories: (1) transport from ganglion cells innervating all receptive elements of the labyrinth, (2) transport from ganglion cells innervating the three semicircular ducts, and (3) transport from cells of the inferior vestibular ganglion innervating the posterior semicircular duct. Transneuronal transport of isotope was observed in secondary vestibular fibers in animals where proline was used and survival exceeded 12 days. Transneuronal labeling of secondary auditory fibers was independent of the [³H]amino acid used, and occurred with survivals of 10 or more days. HRP implanted into the ampulla of the lateral semicircular duct in several animals produced retrograde transport to efferent vestibular and cochlear neurons, but did not result in transganglionic labeling of primary vestibular or auditory fibers.Primary vestibular fibers terminate throughout the superior (SVN) and medial vestibular nuclei (MVN). Within SVN, terminals are most pronounced in its central large-celled portion, but extend into peripheral parts of the nucleus, except for a small medial area near its junction with the oral pole of MVN. Primary projections to MVN are homogenously distributed throughout the nucleus excepting a small circular area of sparse terminals along its ventral margin. Primary vestibular afferents terminate mainly in rostral and caudal portions of the inferior vestibular nucleus (IVN), but do not reach cell group ‘f’. Projections to the lateral vestibular nucleus (LVN) are restricted to its ventral part. Primary projections to the accessory vestibular nuclei reach the interstitial nucleus of the vestibular nerve (NIVN) and cell group ‘y’. Fibers project beyond the vestibular nuclei (VN) to terminate ipsilaterally in the accessory cuneate nucleus (ACN), the subtrigeminal lateral reticular nucleus (SLRN), and well-defined portions of the reticular formation (RF). Projections to SVN and MVN are derived primarily from ganglion cells innervating the semicircular ducts, while projections to caudal IVN, cell group ‘y’ and ACN are related mainly to macular portions of the vestibular ganglion. NIVN receives both macular and duct afferents. Posterior duct afferents terminate in medial portions of SVN, in rostrolateral portions of MVN, and in rostral IVN.Transneuronal transport of isotope increases the volume of terminal label in the ipsilateral VN, but not in dorsal LVN, or cell groups ‘f’ or ‘x’. The quality of transneuronal transport in secondary vestibular fibers is dependent upon: (1) survival time, (2) proximity to the VN, and (3) the excitatory or inhibitory nature of the projection.Primary vestibulocerebellar fibers terminate heavily in the ipsilateral nodulus and ventral uvula. Lesser projections reach the flocculus, deep folia of vermal lobules V and VI, and the lingula. Primary vestibulocerebellar projections terminate as mossy fiber rosettes in the granular layer of these cortical areas. No primary vestibular fibers terminate in the primate fastigial nuclei.     

Central trajectories of type II spiral ganglion neurons

Previous attempts to trace the central pathways of the thin axons from type II spiral ganglion neurons have been hampered by technical difficulties such as fading of the reaction product as distance increases from the injection site (Ryugo et al.: Soc. Neurosci. Abstr. 12:779, '86; Brown: J. Comp. Neurol. 260:591-604, '87). By using small rodents (gerbils and mice), which have short auditory nerves, we have succeeded in filling the entire central axon and terminals of type II neurons after peripheral injections of horseradish peroxidase. The general course of the type II fibers within the auditory nerve and cochlear nucleus is similar to that of type I fibers except that terminals from type II neurons are often found in regions of the cochlear nucleus that have high densities of granule cells.     

Posteroventral cochlear nucleus projections to olivocochlear neurons

The presence of ascending auditory inputs from the posteroventral cochlear nucleus (PVCN) to olivocochlear neurons was examined in guinea pig by using the combination Phaseolus vulgaris-leucoagglutinin (PHA-L) anterograde and horseradish peroxidase (HRP) retrograde tract-tracing technique. By labeling the somata of olivocochlear neurons after injection of HRP into the cochlea and simultaneously labeling terminal endings of PVCN efferent neurons after injection of PHA-L into PVCN, we observed neuronal connections between these two elements within all regions of the superior olivary complex known to contain olivocochlear neurons. These regions include the superior paraolivary nucleus, medial nucleus of the trapezoid body, lateral superior olive, and periolivary regions. All possible projection patterns regarding side of input and output of both large (four combinations) and small (two combinations) olivocochlear neurons were observed. However, the most frequently observed pattern was the PVCN projection to a contralaterally located and contralaterally projecting, large olivocochlear neuron. Thus the most prevalent pattern demonstrated a feedback pathway that crossed the brainstem twice. Additional patterns demonstrated pathways that fed back to the same cochlea as well as pathways that fed forward to the opposite cochlea.     

Projections of the second cervical dorsal root ganglion to the cochlear nucleus in rats

Physiological, anatomical, and clinical data have demonstrated interactions between somatosensory and auditory brainstem structures. Spinal nerve projections influence auditory responses, although the nature of the pathway(s) is not known. To address this issue, we injected biotinylated dextran amine into the cochlear nucleus or dorsal root ganglion (DRG) at the second cervical segment (C2). Cochlear nucleus injections retrogradely labeled small ganglion cells in C2 DRG. C2 DRG injections produced anterograde labeling in the external cuneate nucleus, cuneate nucleus, nucleus X, central cervical nucleus, dorsal horn of upper cervical spinal segments, and cochlear nucleus. The terminal field in the cochlear nucleus was concentrated in the subpeduncular corner and lamina of the granule cell domain, where endings of various size and shapes appeared. Examination under an electron microscope revealed that the C2 DRG terminals contained numerous round synaptic vesicles and formed asymmetric synapses, implying depolarizing influences on the target cell. Labeled endings synapsed with the stalk of the primary dendrite of unipolar brush cells, distal dendrites of presumptive granule cells, and endings containing pleomorphic synaptic vesicles. These primary somatosensory projections contribute to circuits that are hypothesized to mediate integrative functions of hearing.     

Central projections of intracellularly labeled auditory nerve fibers in cats: morphometric correlations with physiological properties

The central arborizations and endings of type I spiral ganglion neurons were labeled with intracellular injections of horseradish peroxidase (HRP) after their characteristic frequency (CF) and spontaneous discharge rate (SR) were physiologically determined. A fiber-by-fiber analysis was conducted and the morphological data compared with the fiber's response properties. The total number of branch points was correlated with total fiber length, a relationship that remained relatively constant when analyzing the ascending and descending branches together or separately. On the other hand, the ascending branches of four out of five fibers having CFs below 0.5 kHz bifurcated and gave rise to a pair of terminal endbulbs of Held. Low- and medium-SR fibers gave rise to more endings than did high-SR fibers, especially on the ascending branch. This difference was accounted for by small endings, a category composed of terminal boutons, string endings, and small complex endings. The categories of modified endbulbs, and endbulbs of Held did not vary in number with respect to fiber SR. The mean area of each ending type within the small ending category was statistically smaller for low- and medium-SR fibers than for high-SR fibers, whereas the mean area of modified endbulbs and endbulbs of Held was not correlated with fiber SR. Total ending area per fiber appeared independent of either CF or SR. These results are discussed in relation to issues of conservation of axon arborizations and terminals, and convergence of input from the different SR groups.     

Topographic organization of the central projections of the spiral ganglion in cats

The morphological organization of inputs from restricted sectors of the cat cochlear spiral ganglion into the cochlear nucleus was studied by making focal extracellular injections of horseradish peroxidase (HRP) into the spiral ganglion. Injections resulted in Golgi-like labeling of a small cluster of spiral ganglion cells and their peripheral and central axons. Large injections involved most of the cells within Rosenthal's canal in sectors of the spiral ganglion innervating greater than or equal to 1 mm of the basilar membrane and resulted in narrow, complete laminae of labeled axons and preterminal fields within each cochlear nucleus subdivision. The positions of these bands were consistent with the "isofrequency laminae" appropriate for the frequencies represented at the injection sites, with high frequency laminae situated more dorsally, and lower frequencies progressively more ventral. A discrete projection to the small cell cap area was observed that was discontinuous with the main projection laminae in the ventral cochlear nuclei (VCN). In the dorsal cochlear nucleus, projecting fibers and terminals were excluded from the molecular cell layer. No labeled fibers entered the granule cell areas. In contrast to larger injections, very small HRP deposits labeled only part of an isofrequency lamina. Specifically, injections restricted to the scala tympani aspect of the spiral ganglion labeled only the lateral part of VCN isofrequency laminae, whereas injections limited to the scala vestibuli aspect of the ganglion labeled the medial aspect of the isofrequency planes. Thus these data indicate a previously unrecognized topographic representation of the vertical dimension of the spiral ganglion across VCN isofrequency laminae. Some possible functional implications of this projection organization are discussed.     

CNS projections to the pterygopalatine parasympathetic preganglionic neurons in the rat: a retrograde transneuronal viral cell body labeling study.

The retrograde transneuronal viral cell body labeling method was used to study the CNS nuclei that innervate the parasympathetic preganglionic neurons which project to the pterygopalatine ganglion. Small injections of a suspension of pseudorabies virus (PRV) were made in the pterygopalatine ganglion of rats and after 4 days their brains wer e processed for immunohistochemical detection of PRV. Some of the tissues were stained with a dual immunofluoresence method that permitted the visualization of PRV and neurotransmitter enzyme or serotonin immunoreactivity in the same cell. Retrograde cell body labeling was detected in the ipsilateral ventrolateral medulla oblongata in the region that has been termed the superior salivatory nucleus. This area was the same region that was retrogradely labeled after Fluoro-Gold dye injections in the pterygopalatine ganglion. Retrograde transneuronally infected cell bodies that provide putative afferent inputs to the pytergopalatine parasympathetic preganglionic neurons were mapped throughout the brain. In the medulla oblongata, transneuronally labeled neurons were seen in the nucleus tractus solitarii, dorsomedial part of the spinal trigeminal nucleus and gigantocellular reticular nucleus. In most experiments, some A1 catecholamine cells and serotonin neurons of the raphe magnus, raphe pallidus, raphe obscurus, and parapyramidal nuclei were labeled. In the pons, labeled cells were found in the parabrachial nucleus. A5 catecholamine cell group, and non-catecholamine part of the subcoeruleus region. In the midbrain, cell body labeling was located in the central gray matter and retrorubral field. In the diencephalon, labeling was found mainly in the hypothalamus. The areas included the lateral hypothalamic area, lateral preoptic area, dorsomedial and paraventricular hypothalamic nuclei, and ventral zona incerta. Contralateral second order cell body labeling was seen in the tuberomammillary nucleus of the hypothalamus. Some of these cells were histidine decarboxylase-immunoreactive. In the forebrain, the bed nucleus of the stria terminalis, substantia innominata, and an area of the cerebral cortex called the amygdalopiriform transition zone were labeled.     

Transport of radioactivity from primary auditory neurons beyond the cochlear nuclei.

Transneuronal degeneration in secondary brainstem auditory nuclei has been described following cochlear lesions, and Marchi studies have revealed degeneration in the trapezoid body after such lesions. These findings support the view of some early neuroanatomists that primary auditory afferents project beyond the cochlear nuclei. Silver impregnation studies of degenerating cochlear afferents have failed to support this claim, leaving the question unresolved. Using [3H] amino acids auditory projections in the monkey were traced autoradiographically from labeled ganglion cells: (1) in all portions of the spiral ganglion, (2) in apical and middle turns of the cochlea and (3) primarily in the basal turn of the cochlea. Labeling of the entire spiral ganglion resulted in profuse transport of isotope to parts of all cochlear nuclei [anteroventral (AVCN), posteroventral (PVCN), and dorsal (DCN) cochlear nuclei]. Labeled axons entering the trapezoid body (TB) projected ipsilaterally to: (1) the lateral trapezoid nuclei (LTN), (2) the lateral superior olivary nucleus (LSO), (3) the dorsal dendritic zone of the medial superior olivary nucleus (MSO), and (4) the pericollicular or cortical nucleus of the inferior colliculus (IC). Auditory fibers crossing the midline in the TB projected contralaterally to: (1) the medial trapezoid nucleus (MTN), (2) the ventral dendritic zone of the MSO, (3) the ventral nucleus of the lateral lemniscus (VNLL), and (4) the central nucleus of the inferior colliculus (CNIC). Selective labeling of cells in the apical and middle ganglionic turns ofthe cochlea resulted in a similar pattern of centripetal transport. Selective [3H] uptake primarily by ganglion cells in the basal turn of the cochlea resulted in: (1) labeling of restricted parts of all cochlear nuclei, and (2) a considerable reduction of isotope transported beyond the cochlear nuclei. Labeled fibers in the TB were distributed sparsely in the ipsilateral LTN, LSO and dorsal dendritic zone of the MSO. Fibers in the TB were not labeled contralaterally in these animals. Interpretation of these data considers the possibility of transneuronal isotope transport. The pattern of isotope transported beyond the cochlear nuclei does not correspond to the secondary projections of any individual cochlear nucleus, although it bears some resemblance to that of the AVCN. The current study suggests that some primary auditory afferents project beyond the cochlear nuclei.     

Transneuronal transport of WGA-HRP in immature rat visual pathways

Wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) was used to study transneuronal transport in the developing rat visual pathways. Intraocular injections of WGA-HRP were made in neonatal albino rat pups at different ages from the day of birth, postnatal day 0 (P0), to one month of age. Transneuronal labeling in geniculostriate fibers and in tectoparabigeminal terminals was observed as early as P1 and showed little change with eye-opening. However, at early ages, consistent transneuronal labeling was found to require injection of up to 4 times the amount of tracer (0.18 mg WGA-HRP) as adults (0.04 mg WGA-HRP), delivered in two injections. Control injections of HRP alone produced heavy anterograde labeling at all ages, without requiring increased injections. The results suggest that transneuronal transport precedes synaptic transmission, and may illustrate a mechanism for exchanging molecules between neurons. One explanation for the requirement of increased tracer is that axonal and/or transneuronal transport of WGA-HRP may be selectively limited to certain cells in the postnatal retina.     

Cochlear-nucleus branches of thick (medial) olivocochlear fibers in the mouse: a cochleotopic projection

Olivocochlear neurons have somata in the superior olivary complex and provide an efferent innervation to the cochlea. One subgroup of olivocochlear neurons, medial olivocochlear neurons, sends fibers to innervate the cochlear outer hair cells. En route to the cochlea, medial olivocochlear fibers give off branches to the ventral cochlear nucleus, the first auditory center of the brain. This study examines the cochlear-nucleus branches of medial olivocochlear fibers, comparing those from fibers that innervate the cochlear base with those from fibers that innervate the cochlear apex. Basal fibers give off dorsal branches to the granule cell lamina and ventral branches to the auditory nerve root. Apical fibers give off few dorsal branches but many ventral branches that terminate rostrally to the nerve root. This cochleotopic mapping of medial olivocochlear branches corresponds in a general way to that of afferent fibers. Unlike afferent fibers, however, the branches terminate primarily along the edges of the cochlear nucleus. In the mouse, the particular edges of termination are (1) the medial border of the ventral cochlear nucleus where it meets the underlying vestibular nerve root, and (2) the border between the ventral cochlear nucleus and the granule cell lamina. Neurons and dendrites of these border regions may thus integrate efferent and afferent information in a frequency-specific manner.