Selective retrograde labeling of neurons of the cat vestibular ganglion with [3H]D-aspartate

D-[2,3-3H]Aspartate [( 3H]D-Asp) was injected in the cat vestibular nuclei. Labeling patterns resulting from retrograde axonal transport by the vestibular nerve fibers were observed in the vestibular ganglion neurons and also in the nerve fibers. The selectivity of such labeling, related to the neurotransmitter's specificity, is strongly indicated.     

Chemical anatomy of excitatory endings in the dorsal cochlear nucleus of the rat: Differential synaptic distribution of aspartate aminotransferase,glutamate, and vesicular zinc

In order to identify cytochemical traits relevant to understanding excitatory neurotransmission in brainstem auditory nuclei, we have analyzed in the dorsal cochlear nucleus the synaptic distribution of aspartate aminotransferase, glutamate, and vesicular zinc, three molecules probably involved in different steps of excitatory glutamatergic signaling. High levels of glutamate immunolabeling were found in three classes of synaptic endings in the dorsal cochlear nucleus, as determined by quantitation of immunogold labeling. The first type included auditory nerve endings, the second were granule cell endings in the molecular layer, and the third very large endings, better described as “mossy.” This finding points to a neurotransmitter role for glutamate in at least three synaptic populations in the dorsal cochlear nucleus. The same three types of endings enriched in glutamate immunoreactivity also contained histochemically detectable levels of aspartate aminotransferase activity, suggesting that this enzyme may be involved in the synaptic handling of glutamate in excitatory endings in the dorsal cochlear nucleus. There was also extrasynaptic localization of the enzyme. Zinc ions were localized exclusively in granule cell endings, as determined by a Danscher-selenite method, suggesting that this ion is involved in the operation of granule cell synapses in the dorsal cochlear nucleus. J. Comp. Neurol. 399:341–358, 1998. © 1998 Wiley-Liss, Inc.     

Afferent fibers in the hypoglossal nerve: a horseradish peroxidase study in the cat

The existence of afferent fibers in the cat hypoglossal nerve was studied by transganglionic transport of horseradish peroxidase (HRP). Injections of wheat germ agglutinin-conjugated HRP (WGA-HRP) into the hypoglossal nerve resulted in some retrograde labeling of cell bodies within the superior ganglia of the ipsilateral glossopharyngeal and vagal nerves. A few labeled cell bodies were also present ipsilaterally within the inferior ganglion of the vagal nerve and the spinal ganglion of the C1 segment. Some of the labeled glossopharyngeal and vagal fibers reached the nucleus of the solitary tract by crossing the dorsal portion of the spinal trigeminal tract. Others distributed to the spinal trigeminal nucleus pars interpolaris and to the ventrolateral part of the medial cuneate nucleus by descending through the dorsal portion of the spinal trigeminal tract. In the spinal cord these descending fibers, intermingling with labeled dorsal root fibers, distributed to laminae I, IV-V and VII-VIII of the C1 and C2 segments. Additional HRP experiments revealed that the fibers in laminae VII-VIII originate mainly from dorsal root of the C1 segment.     

Descending projections from the superior olivary complex to the cochlear nucleus of the cat

Subdivisions of the cochlear nuclear complex give rise to a number of discrete projections to certain cell groups of the superior olivary complex and also received substantial descending projections from the periolivary nuclei. In the present study, we sought to determine by means of retrograde transport of horseradish peroxidase (HRP), and anterograde transport of radiolabeled protein, if the periolivary nuclei give rise to discrete projections to the various subdivisions of the cochlear nuclear complex. Following medium to large injections of HRP into the cochlear nucleus, irrespective of location, labeled cells were found in all periolivary nuclei bilaterally. In every case more than 40% of the labeled cells were found in the lateral nucleus of the trapezoid body on the same side and the ventral nucleus of the trapezoid body of both sides. Other periolivary nuclei contributing more than 5% of the total number of cells in individual cases were the contralateral lateral nucleus of the trapezoid body and the ipsilateral anterolateral and dorsal periolivary nuclei. Injections of tritiated leucine into periolivary nuclei gave rise to axonal labeling to the trapezoid body and the dorsal acoustic stria, usually bilaterally, and to terminal labeling that was widely distributed within the cochlear nuclear complex. In several cases with small injections, particularly in the lateral nucleus of the trapezoid body, the projections from the periolivary nuclei to the anteroventral and dorsal cochlear nuclei connected areas described as having similar best-frequency representation. The autoradiographic data corroborated the main results from the HRP experiments and provided additional information permitting these conclusions: the projections from the periolivary nuclei to the cochlear nuclear complex are organized tonotopically, at least in part; each periolivary nucleus (and perhaps individual cells), projects widely throughout the cochlear nuclear complex; the pattern of termination of projections from different periolivary nuclei to a given region of the cochlear nuclear complex are similar, as seen in autoradiograms, and the lateral and dorsal periolivary nuclei project mainly ipsilaterally, while the medial periolivary nuclei project bilaterally with a contralateral bias. The magnitude of these projections and their widespread distribution within the cochlear nuclear complex would suggest an important role for the descending projections in the normal functioning of the cochlear nucleus.     

Primary projections and efferent cells of the VIIIth cranial nerve in the monitor lizard, Varanus exanthematicus

In order to describe the central relations of both the afferent and efferent components of the VIIIth cranial nerve in one reptile, the methods of anterograde and retrograde axonal transport and anterograde degeneration were used to study the vestibular and cochlear projections and the efferent system of this nerve in Varanus exanthematicus. On the basis of cresyl violet and Klüver-Barrera staining, five vestibular nuclei, four cochlear nuclei, and two clusters of small cells which could not be designated as strictly auditory or vestibular are distinguished. The vestibular nuclei include the nucleus dorsolateralis, nucleus ventrolateralis, nucleus tangentialis, nucleus ventromedialis, and nucleus descendens. The well-developed cochlear nuclear complex includes the nucleus angularis, nuclei magnocellulares medialis and lateralis, and nucleus laminaris. The two cell clusters are located dorsolaterally in the brainstem just ventrolateral to the acoustic tubercle. The primary afferent vestibular fibers coursing in the anterior VIIIth nerve root distribute to the ventral portions of all vestibular nuclei except nucleus ventromedialis, whereas the fibers coursing in the posterior root project to the dorsal portions of these nuclei. In nucleus ventromedialis fibers of both roots do not segregate into ventral and dorsal portions. Other targets of the vestibular fibers are the two cell clusters, the granular layer of the ipsilateral cerebellum, the reticular formation, and the descending trigeminal tract and its nucleus. The primary cochlear fibers coursing in the posterior root terminate in nucleus angularis, nuclei magnocellulares medialis and lateralis, and the inner cell strand of nucleus laminaris. The efferent system is, ipsi- and contralaterally in the brainstem, composed of ventral and dorsal cell groups that extend from the level of the principal abducens nucleus caudally where they overlap with the facial motor nucleus. The fibers, which originate from the contralaterally located efferent cells, course beneath the IVth ventricle to exit the brainstem on the ipsilateral side.     

Pathways connecting the right and left cochlear nuclei

Connections between the right and left cochlear nuclei were studied with retrograde and anterograde axonal transport techniques. Large, multipolar neurons in the anterior and posterior divisions of the anteroventral cochlear nucleus and in the posteroventral cochlear nucleus project to the ventral and dorsal cochlear nuclei on the opposite side. In addition, giant cells in the deep layers of the dorsal cochlear nucleus project to the contralateral posteroventral cochlear nucleus and possibly also to the contralateral dorsal cochlear nucleus. The pattern of terminal distribution of the crossed connections was determined by using the anterograde axonal transport of horseradish peroxidase-labelled wheat germ lectin. Although no part of the cochlear nuclear complex is completely free of anterograde label, the densest labelling is found in the anterior division of the anteroventral cochlear nucleus, throughout the posteroventral cochlear nucleus (where it is closely associated with cell bodies), and in the fusiform and superficial layers of the dorsal cochlear nucleus. These direct synaptic connections from one cochlear nucleus to the other could play a significant role in processes that depend on binaural interactions within the central nervous system.     

Distribution and origin of serotoninergic afferents to guinea pig cochlear nucleus

The distribution of serotoninergic fibers in the guinea pig cochlear nucleus was studied with serotonin immunohistochemistry. In addition, the origin of the serotoninergic fibers was determined by combining the retrograde transport of wheat germ agglutinin-apohorseradish peroxidase (gold conjugated) with serotonin immunohistochemistry. Immunoreactivity was present in varicose and nonvaricose fibers that were unevenly distributed throughout the cochlear nucleus. The fibers were most prominent in the superficial layers of the dorsal cochlear nucleus and the anterior spherical cell area of the anteroventral cochlear nucleus. Although less prominent, serotonin-positive fibers were also present in the remaining part of the anteroventral cochlear nucleus and the posteroventral cochlear nucleus. A few positive fibers were present in the auditory nerve root and the dorsal and intermediate acoustic stiae. Double-labeled cells were found throughout the rostral- caudal extent of the serotoninergic system from the caudal linear nucleus to the nucleus raphe pallidus. However, most were confined to the dorsal (52%) and median (18%) raphe nuclei. Some serotoninergic cell groups contained retrogradely labeled cells that were not serotonin immunoreactive, indicating nonauditory afferents to cochlear nucleus containing other neurotransmitter substances. Serotonin may tonically modulate auditory processing within the cochlear nucleus as well as influence certain ascending auditory pathways. Most of the serotonin in the cochlear nucleus comes from superior raphe nuclei that also project to basal ganglia motor systems and limbic strctures. Therefore, the effect of serotonin on the cochlear nucleus may be related to level of arousal or behavioral state. © 1995 Willy-Liss, Inc.     

Direct projections from the dorsal column nuclei and the spinal trigeminal nuclei to the cochlear nuclei in the cat

A retrograde and anterograde wheat germ agglutinated horseradish peroxidase WGA-HRP study in the cat indicated that some neurons in the dorsal column nuclei and the interpolar and caudal spinal trigeminal nuclei send fibers to the dorsal and ventral cochlear nuclei; to the pyramidal cell layer of the dorsal cochlear nucleus and to the cochlear granule cell domain, bilaterally with an ipsilateral dominance.     

Peripheral nerve autografts to the rat spinal cord: Studies with axonal tracing methods

In young adult female rats, autologous sciatic nerve segments were transplanted to the thoracic region of the spinal cord. The grafts became well innervated but led to no obvious functional improvement. The origin and termination of axons in the grafts was studied by retrograde neuronal labeling with horseradish peroxidase (HRP) and radioautographic axonal tracing. Studies with HRP indicated that some axons in the grafts originated from intrinsic CNS neurons with their cell bodies in nearby segments of the spinal cord and that others arose from dorsal root ganglia at the level of the grafts and at least 7 segments distal to them. After tritiated amino acids were injected into lumbar dorsal root ganglia, labeled axons could be followed into the grafts but not into the rostral spinal cord stumps. Together with other experimental observations, these results demonstrate a correlation between success or failure of elongation of dorsal root fibers and peripheral or central ensheathment at the axonal tip. The corticospinal tract was studied both with radioautography and retrograde axonal transport of HRP but no extension of its axons into peripheral nerve grafts was detected under these experimental conditions. The findings implicate both neuroglial and axonal factors in the feeble regenerative response usually seen after injury to the spinal cord.     

Observations on the orthograde and retrograde transport of horseradish peroxidase in the cat.

Horseradish peroxidase (HRP) was applied to the precruciate cortex of the cat. The peroxidase was either applied directly to the surface or injected by glass micropipettes in single or multiple injections. Retrograde as well orthograde transport was observed by light and electron microscopy in different areas: in the thalamus: the nucleus ventralis lateralis and the nucleus centrum medianum, in the red nucleus and in the spinal cord. The distribution of HRP within the nervous elements of the nuclei: axon terminals and/or perikarya and dendrites, was observed by light and electron microscopy. HRP was present in dendrites and perikarya, indicating retrograde transport, in experiments where damage to the injected nervous tissue could be suspected. However HRP appeared restricted to the nerve endings, indicating orthograde transport, when the damage seemed to be minimal. This effect of injury on retrograde transport of HRP was further confirmed by experiments using bilateral HRP injections and unilateral lesions of the spinal cord to label rubro-spinal cells. Thus retrograde transport of HRP could be induced by lesion in systems where it does not occur normally. The normal intra-axonal transport of HRP appears to be from the perikaryon to the nerve endings. This orthograde transport can be seen under the light microscope and can be confused with faint retrograde labeling.     

Afferent projections to the central nucleus of the inferior colliculus in the rat

The afferent projections to the inferior colliculus of the rat were studied using the method of retrograde transport of horseradish peroxidase (HRP).Following large injections of HRP into the central nucleus, cells within the cochlear nuclei, superior olivary complex and auditory cortex were stained. Within the contralateral dorsal cochlear nucleus, fusiform cells were heavily labeled. Giant cells were also labeled in deeper layers. In the contralateral ventral cochlear nucleus, virtually all major cell types were labeled, with some types being labeled in greater numbers than others. Octopus cells of posteroventral division of ventral cochlear nucleus (PVCN) were never labeled. HRP-positive cells were found in ipsilateral and contralateral lateral superior olivary nucleus (LSO), ipsilateral medial superior olivary nucleus (MSO), ipsilateral and contralateral lateral nucleus of the trapezoid body (LTB), ipsilateral ventral nucleus of the trapezoid body (VTB), and ipsilateral superior paraolivary nucleus (SPN). Pyramidal cells of layer V of auditory cortex were heavily labeled.Small injections of HRP into the central nucleus resulted in labeled cells within restricted regions of the cochlear nuclei, superior olivary complex and auditory cortex. Injections into dorsal regions of the central nucleus resulted in cells labeled in ventral regions of the dorsal and ventral cochlear nuclei, and in lateral regions of LSO. These regions contain neurons which are considered to have low best frequencies. Injections placed in more ventral regions of the central nucleus led to labeling of cells in more dorsal regions of the cochlear nuclei and more medial regions of LSO in agreement with the tonotopical progressions within these structures.     

The central distribution of the cervical vagus nerve and gastric afferent and efferent projections in the rat

The medullary distribution of afferent fibers and cells of origin of the cervical vagal trunk and of the vagal innervation of the stomach have been studied using the anterograde and retrograde transport of horseradish peroxidase (HRP). Injections of HRP were made into the cervical vagus nerve, the stomach wall, the proximal small intestine, or the peritoneal cavity. Two to four days following the injections, the rats were perfused and the medullae oblongatae and nodose ganglia were processed using the tetramethyl benzidine method. Cervical vagus nerve injections of HRP resulted in heavy anterograde labeling in the ipsilateral nucleus of the tractus solitarius (NTS) and the commissural nucleus. Lighter labeling was seen in these regions on the contralateral side, but did not extend as far rostrally in the NTS. Labeling was also seen in the area postrema. Retrogade labeling of somata was present in the ipsilateral side in the nodose ganglion, throughout the whole extent of the dorsal motor nucleus of the vagus, much of the nucleus ambiguus and in rostral levels of the cervical spinal cord. After stomach injections, labeling indicative of afferent fibers was observed bilaterally in the dorsomedial and medial portions of the NTS and in the commissural nucleus. Labeled efferent fibres arose from neurons in the dorsal motor nucleus of the vagus, nucleus ambiguus and the cervical spinal cord. Retrogradely labeled somata were found bilaterally, throughout the rostrocaudal length of the dorsal motor nucleus in all cases with stomach injections. In some, but not all cases, labeled somata were seen bilaterally in compact areas within the nucleus ambiguus, particularly rostrally. Control injections of HRP into the intestinal wall and peritoneal cavity indicated that the stomach was the primary source of afferent and efferent labeling in the medulla following subdiaphragmatic injections.     

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.     

Selective retrograde labeling of neurons of the cat vestibular ganglion with [H]d-aspartate

d-[2,3-³H]Aspartate ([³H]d-Asp) was injected in the cat vestibular nuclei. Labeling patterns resulting from retrograde axonal transport by the vestibular nerve fibers were observed in the vestibular ganglion neurons and also in the nerve fibers. The selectivity of such labeling, related to the neurotransmitter's specificity, is strongly indicated.     

Brain stem afferents to the fastigial nucleus in the cat demonstrated by transport of horseradish peroxidase.

Although retrograde and anterograde degeneration studies have provided important information concerning brain stem afferents to the fastigal nucleus (FN), these data may be incomplete and should be confirmed by axonal transport methods. Attempts were made to inject horseradish peroxidase (HRP) unilaterally into the FN in a series of adult cats. Animals were perfused with dextran and a fixative solution of paraformaldehyde and glutaraldehyde in 0.1 M phospate buffer. Representative sections were treated by the Graham and Karnovsky ('66) method. Selective HRP injections in one FN resulted in retrograde transport of the marker to Purkinje cells of the ipsilateral vermis and distinctive appendages of the contralateral medial accessory olivary (MAO) nucleus (nucleus beta and the dorso-medial cell column). Retrograde transport of the label was found bilaterally in cells of the medial (MVN) and inferior (IVN) vestibular nuclei, in cell group x and in the nucleus prepositus (PP). Labeled vestibular neurons, most numerous in MVN, were identified in dorsal, caudal and lateral regions, with a slight ipsilateral preponderance. Only a few neurons in caudal, dorsal and lateral regions of the IVN were labeled and none of these included cells of group f. Labeled cells in the caudal third of PP were greatest ipsilaterally. Rostral and caudal injections of FN labeled smaller numbers of cells in MVN, IVN, cell group x and PP. HRP injections of FN and portions of lobules VIII and IX resulted in bilateral retrograde labeling of larger numbers of cells in MVN, IVN and cell group x, and ipsilateral labeling of cells in group y and the interstitial nucleus of the vestibular nerve. Injections of HRP into basal folia of lobules V and VI resulted in retrograde transport of the marker to cells of the medial and dorsal accessory olivary nuclei contralaterally, and to cells of the ipsilateral accessory cuneate nucleus. Transport of label injected into portions of the pyramis was detected in parts of the contralateral MAO and bilaterally in parts of the pontine and reticulotegmental nuclei. This study suggests that the principal afferents of the fastigial nucleus arise from: (1) Purkinje cells of the ipsilateral vermis, (2) restricted portions of the contralateral MAO (nucleus beta and dorsomedial cell column), (3) portions of the MVN and IVN (bilaterally) and (4) caudal parts of the PP. Secondary vestibular inputs to the fastigial nucleus probably are relayed mainly by Purkinje cells in the cerebellar cortex.     

Transneuronal labeling of cochlear nucleus neurons by HRP-labeled auditory nerve fibers and olivocochlear branches in mice.

Auditory nerve fibers were labeled by extracellular injections of horseradish peroxidase into the spiral ganglion in mice. The labeled fibers were traced in an anterograde direction through the auditory nerve into the cochlear nucleus. In almost half of the injections, the labeled endings of auditory nerve fibers contacted cochlear nucleus neurons that were also labeled with horseradish peroxidase and were presumably transneuronally labeled. Only darkly labeled endings were associated with transneuronally labeled neurons, but not all darkly labeled endings had targets that were transneuronally labeled. Transneuronal labeling occurred almost exclusively in the ventral cochlear nucleus, often between endbulbs and bushy cells. Both "modified" endbulbs and the larger endbulbs of Held transneuronally labeled the bushy cells that they contacted. At the ultrastructural level, transneuronal labeling was evident as a darkening of ribosomes and the membrane surfaces of mitochondria, endoplasmic reticulum, and the nucleus. Transneuronal labeling occurred rarely in octopus, small, and stellate cells, and in neurons of the dorsal cochlear nucleus. Spiral ganglion injections also label olivocochlear fibers, efferent fibers that pass through the ganglion en route to the hair cells. These fibers give off branches to the cochlear nucleus that were rarely associated with transneuronal labeling. In eight instances, the targets of olivocochlear branches were stellate cells or small cells. We suggest that in our mouse preparation, horseradish peroxidase is effective as a transneuronal marker because the short distance from injection site to the cochlear nucleus results in a high concentration of horseradish peroxidase in the endings of the auditory nerve fibers.     

Central projections of cervical primary afferent fibers in the guinea pig: an HRP and WGA/HRP tracer study

The central course and the projections of the first and the second cervical dorsal root ganglia and of suboccipital muscle primary afferent fibers in the guinea pig were studied by means of anterograde transport of wheat germ agglutinin conjugated to horseradish peroxidase (WGA/HRP) or aqueous solution of horseradish peroxidase (HRP). Injections of WGA/HRP into the second cervical dorsal root ganglion produced labeling in the dorsal and ventral horns. Within the spinal cord, the largest amount of HRP reaction product was found within the lateral third of the substantia gelatinosa and within the central cervical nucleus. The main area of termination in the medulla was the external cuneate nucleus. However, HRP reaction product was also found within the medial and inferior vestibular nuclei, cell group x, the perihypoglossal nuclei, the nucleus of the solitary tract, and the nucleus of the spinal trigeminal tract. Descending fibers could be detected as caudal as spinal segment T5. Injections of WGA/HRP into the first cervical dorsal root ganglion produced heavy terminal label within the central cervical nucleus but not within the substantia gelatinosa. Again, the external cuneate nucleus was the main area of termination within the medulla. Label could not be observed within the vestibular nuclear complex or within the spinal trigeminal nucleus. Injections of aqueous HRP into the suboccipital muscles produced heavy transganglionic label within the central cervical nucleus, whereas the substantia gelatinosa totally lacked terminal label. Ascending proprioceptive fibers reached the external cuneate nucleus and group x. Scanty projections could be detected within the vestibular nuclei as well as within the perihypoglossal nuclei except for the nucleus prepositus hypoglossi. Label was absent in the spinal trigeminal nucleus.     

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.     

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.     

Neurotransmitter-specific uptake and retrograde transport of [3H]glycine from the inferior colliculus by ipsilateral projections of the superior olivary complex and nuclei of the lateral lemniscus.

Neurotransmitter-specific uptake and retrograde axonal transport of [3H]glycine were used to identify glycinergic projections to the inferior colliculus in chinchillas and guinea pigs. Six h after injection of [3H]glycine in the inferior colliculus, autoradiographically labeled cells were found ipsilaterally in the ventral nucleus of the lateral lemniscus, the lateral superior olive and the dorsomedial periolivary nucleus. These 3 regions accounted for 95% of the labeled projection neurons, with the remainder scattered elsewhere in the ipsilateral superior olivary complex. No labeled cells were found contralaterally even after survival times as long as 24 h. Retrograde transport of HRP from the inferior colliculus in these same cases confirmed the presence of additional projections that did not accumulate [3H]glycine. These included ipsilateral projections from the medial superior olive and cochlear nucleus and contralateral projections from the inferior colliculus, dorsal nucleus of the lateral lemniscus, lateral superior olive, periolivary nuclei and cochlear nucleus. The results implicate uncrossed projections from the ventral nucleus of the lateral lemniscus, lateral superior olive, and dorsomedial periolivary nucleus as the principal sources of inhibitory glycinergic inputs to the inferior colliculus.