Bilateral cochlear ablation in postnatal rat disrupts development of banded pattern of projections from the dorsal nucleus of the lateral lemniscus to the inferior colliculus.

Axonal projections from the dorsal nucleus of the lateral lemniscus (DNLL) distribute contralaterally in a pattern of banded layers in the central nucleus of the inferior colliculus (IC). The banded pattern of DNLL projections is already in the IC by onset of hearing in postnatal rat pups. Previously, it was shown that unilateral cochlear ablation in neonatal rat pups disrupted the banded pattern in IC for the projections of the DNLL contralateral to the ablation but not those of the DNLL ipsilateral to the ablation. In the present study, bilateral cochlear ablation or sham surgery was performed at postnatal day 9 (P9) after which rat pups were killed at P12 and the brains removed to study axonal projections of the DNLL. A lipophilic carbocyanine dye, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI), was placed in the dorsal tegmental commissure of Probst to label decussating DNLL axons that end in the central nucleus of the contralateral IC. The distribution of labeled fibers across the central nucleus of the IC was analyzed in digital images by comparing the pattern of labeling with a sine model of periodic distribution of banded layers. In the control group, labeled axons formed a regular pattern of dense banded layers in IC. In the bilateral cochlear ablation group, labeled axons in the IC were distributed diffusely and there was little or no regular pattern of dense bands of axonal labeling. The influence of the cochlea on developing auditory circuits possibly mediated by activity-dependent mechanisms is discussed.

Development of banded afferent compartments in the inferior colliculus before onset of hearing in ferrets.

Axonal projections from the lateral superior olivary nuclei (LSO), as well as from the dorsal cochlear nucleus (DCN) and dorsal nucleus of the lateral lemniscus (DNLL), converge in frequency-ordered layers in the central nucleus of the inferior colliculus (IC) where they distribute among different synaptic compartments. A carbocyanine dye, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI), was used as a tracer to study the postnatal development of axonal projections in the ferret IC. The results indicated that projections from all three nuclei are present at birth, but are not segregated into bands. During the postnatal week between approximately postnatal days 4 and 12 (P4-P12), axons from LSO proliferate in IC, become more branched, and segregate into a series of bands composed of densely packed fibers and endings. LSO projections in these afferent bands course parallel to IC layers and are separated by intervening regions with few endings. A modest fit of a sine curve (R2>0.15) to the pattern of spacing of LSO projections in IC indicated that regularly spaced bands are forming by P7. Similarly, banded patterns of DCN and DNLL projections to IC have developed by the end of the first postnatal week. Thus, well before hearing onset in ferret (P28-30), three different afferent projections have segregated into banded compartments along layers in the central nucleus of the ferret IC. Possible mechanisms in circuit development are discussed.

Cochlear ablation in adult ferrets results in changes in insulin-like growth factor-1 and synaptophysin immunostaining in the cochlear nucleus.

Afferent activity modulates synaptic plasticity as well as the levels of activity-dependent molecules such as growth factors. Disruption of this activity due to deafferentation has been shown to result in an altered trophic support and consequently in changes in neuronal excitability and synaptic transmission. In the present study, to test whether lack of cochlear integrity results in changes in insulin-growth factor-1 (IGF-1) and synaptophysin immunostaining in the cochlear nucleus, the first relay structure in the auditory pathway, unilateral cochlear ablations were performed in adult ferrets. Changes in IGF-1 and synaptophysin immunostaining were assessed in the anteroventral (AVCN), posteroventral (PVCN) and dorsal cochlear nucleus (DCN) at 1, 20 and 90 days after deafferentation. An increase in IGF-1 immunostaining within AVCN, PVCN and DCN was observed ipsilaterally at all survival times after cochlear ablation when compared with the contralateral side and unoperated animals. This increase was accompanied by a significant ipsilateral increase in the mean gray level of synaptophysin immunostaining as well as a decrease in the area of synaptophysin immunostaining at 1 and 20 days after the ablation in AVCN, PVCN and DCN compared with the contralateral side and control animals. These changes in synaptophysin immunostaining were no longer present 90 days after cochlear ablation. The present results provide evidence of a persistent upregulation in IGF-1 and a transitory upregulation in synaptophysin levels in the cochlear nucleus that may reflect neuroprotective mechanisms following the loss of trophic support from spiral ganglion neurons.

Unilateral cochlear ablation before hearing onset disrupts the maintenance of dorsal nucleus of the lateral lemniscus projection patterns in the rat inferior colliculus.

During postnatal development, ascending and descending auditory inputs converge to form fibrodendritic layers within the central nucleus of the inferior colliculus (IC). Before the onset of hearing, specific combinations of inputs segregate into bands separated by interband spaces. These bands may define functional zones within the IC. Previous studies in our laboratory have shown that unilateral or bilateral cochlear ablation at postnatal day 2 (P2) disrupts the development of afferent bands from the dorsal nucleus of the lateral lemniscus (DNLL) to the IC. These results suggest that spontaneous activity propagated from the cochlea is required for the segregation of afferent bands within the developing IC. To test if spontaneous activity from the cochlea also may be required to maintain segregated bands of DNLL input, we performed cochlear ablations in rat pups at P9, after DNLL bands already are established. All animals were killed at P12 and glass pins coated with carbocyanine dye, DiI (1,1'-dioctodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate), subsequently were placed in the commissure of Probst to label the crossed projections from both DNLLs. When compared with surgical controls, experimental results showed a similar pattern of DNLL bands in the IC contralateral to the ablated cochlea, but a disruption of DNLL bands in the IC ipsilateral to the cochlear ablation. The present results suggest that cochlear ablation after DNLL bands have formed may affect the maintenance of banded DNLL projections within the central nucleus of the IC.

Quantitative assessment of developing afferent patterns in the cat inferior colliculus revealed with calbindin immunohistochemistry and tract tracing methods.

The central nucleus of the inferior colliculus (CNIC) is comprised of an orderly series of fibrodendritic layers. These layers include integrative circuitry for as many as 13 different ascending auditory pathways, each tonotopically ordered. Calcium-binding proteins, such as calbindin-D28k (CB), may be useful neurochemical markers for specific subsets of afferent input in these layers and their spatial organization that are developmentally regulated. In this study, CB-immunohistochemistry was used to examine 1-42 postnatal-day-old kitten and adult cat CNIC and anterograde tracers were used to label afferent projections from the lateral superior olivary nucleus (LSO) to the CNIC at similar ages. A distinct axonal plexus that is CB-immunopositive is described. This CB-afferent compartment is present at birth and persists throughout the ages examined. Already at birth, the CB-immunostained plexus in kitten CNIC is organized into discrete bands that are approximately 75 microm thick and 500 microm long. In adult CNIC, the periodic banded pattern of CB-immunostained fibers is similar to that in kittens albeit bands are thicker (145 microm) and longer (700 microm). Growth in band thickness in adult cat appears proportional to growth of the IC, whereas length of the dense CB-immunostained bands is somewhat more focused in the central region of fibrodendritic layers. The banded pattern of the CB-immunostained plexus is well correlated with the location and dimension of afferent projections from the LSO in newborn kitten labeled with carbocyanine dye, 1,1'-dioctodecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate and in adult cat labeled with wheat germ agglutinin conjugated with horseradish peroxidase. The results reveal a neurochemical marker for one type of synaptic compartment in CNIC layers, banding, that is organized before hearing onset in kittens, but that may undergo some postnatal pruning.

Unilateral cochlear ablation in adult ferrets results in upregulation in calretinin immunostaining in the central nucleus of the inferior colliculus.

In the present study, unilateral cochlear ablations were performed in adult ferrets in order to determine whether an upregulation of the calretinin immunostained plexus in the central nucleus of the inferior colliculus occurs and if so, what the time course of this upregulation is. Accordingly, the mean gray level and the calretinin-immunostained area of the axonal plexus in the central nucleus of the inferior colliculus were evaluated at 1, 20 and 90 days after cochlear ablation. In unoperated animals, the calretinin-immunostained plexus was bilaterally symmetric. In ablated animals, both the mean gray level and the immunostained area of the plexus increased in the central nucleus of the inferior colliculus contralateral to the lesion compared with both the ipsilateral side and unoperated animals. This upregulation was present 24 h after the ablation and did not change at the two subsequent time points. In a previous study in young ferrets, the immunostained area of the plexus in the central nucleus of the inferior colliculus contralateral to the lesion increased 200% compared with control ferrets [J Comp Neurol 460 (2003) 585], whereas it increased only 33% in adult ferrets. These findings suggest that 1) calretinin upregulation in the contralateral central nucleus of the inferior colliculus following cochlear ablation occurs by 24 h after cochlear ablation and 2) there is an age-related decline in the magnitude of this upregulation after cochlear ablation.

Afferent sources of a lateral midbrain tegmental zone associated with the pinnae in the cat as mapped by retrograde transport of horseradish peroxidase.

A paralemniscal zone in the lateral midbrain tegmentum of the cat has been identified in a possible pathway from the superior colliculus to the facial nucleus that may control pinna movements (Henkel and Edward, '78). Other brainstem afferent projections to this paralemniscal zone have been mapped in the present study using the retrograde horseradish peroxidase tracing method and are discussed in three groups. First, potential sources of auditory afferents were limited mainly to the external nucleus of the inferior colliculus, the nucleus sagulum, and the dorsomedial periolivary cell group. Labeled cells in other superior olivary regions and the dorsal cochlear nucleus were apparently related to uptake of horseradish peroxidase from the axons of the lateral lemniscus. Second, afferents from several premotor regions involved in aspects of gaze control were identified. These were mainly from the nucleus prepositus hypoglossi and adjacent pontine reticular formation, but also included projections from the medial vestibular an abducens nuclei and possibly subthalamic regions such as the zona incerta and fields of Forel. Third, a relatively large group of midbrain afferents was closely related to the origin of the collicular projection to the paralemniscal zone. This group consisted of labeled cells in the periaqueductal gray matter, nucleus cuneiformis, and pretectum. The relatively sparse labeling in auditory regions that projected to the paralemniscal zone seems to indicate that the sensorimotor integration necessary to guide pinna movements does not take place primarily in the lateral midbrain tegmentum. The interaction of gaze-related sources with the pinna-related pathway is also discussed.

Axonal morphology in fibrodendritic laminae of the dorsal nucleus of the lateral lemniscus: afferent projections from the medial superior olivary nucleus.

The distribution and morphology of axons projecting from the medial superior olivary nucleus to the dorsal nucleus of the lateral lemniscus were studied in the adult cat. Injections of Phaseolus vulgaris-leucoagglutinin, biocytin, or dextran-rhodamine in the medial superior olivary nucleus labeled axons that ascended in the lateral lemniscus. Before entering the inferior colliculus, collateral branches of these labeled axons ended in the dorsal nucleus of the lateral lemniscus in thin, horizontal bands forming laminae that extended throughout the rostral-caudal length of the dorsal nucleus of the lateral lemniscus. A dorsal-ventral topography was apparent in the position of the lamina with respect to the injection site, but no relation between the rostral-caudal location of labeled endings and the injection site was observed. There was a divergent pattern of connections within the horizontal laminae rather than a point-to-point organization. The terminal branches of the collateral axons exhibited round or oval boutons en passant and terminaux. Individual arbors reconstructed from serial sections distributed varicosities in circumscribed domains that were only a subcomponent of the area of the afferent laminae in which they were distributed. The spatial relationships of axonal domains of several axons labeled from a single injection in the medial superior olivary nucleus suggest a mosaic pattern in the laminar connections with the dorsal nucleus of the lateral lemniscus.

The vestibular complex of the American opossum, Didelphis virginiana. I. Conformation, cytoarchitecture and primary vestibular input.

Degeneration experiments reveal that the vestibular nerve of the adult opossum distributes to an extensive and diverse area of the brainstem and, as in placental mammals, the traditionally named nuclei do not receive a uniform distribution of primary afferent fibers. Nevertheless, such nuclei as well as other nearby cell groups have been considered as vestibular since they share other mutual connections. Except for subgroups "l", "g" and the nucleus supravestibularis all of the vestibular subdivisions generally recognized in more specialized placentals (e.g., the cat) have been identified in the marsupial opossum. Each of the vestibular nuclei and "extra-nuclear" cell stations are described as to their boundaries, cytoarchitecture and dendritic domain. Whenever possible we have employed a terminology consistent with that used for the cat. In several instances we have used results from experimental degeneration material to identify nuclear boundaries and divisions which are not apparent in routine Nissl, Golgi or silver preparations.

The superior colliculus control of pinna movements in the cat: possible anatomical connections.

Possible anatomical pathways mediating superior colliculus control of pinna movements were determined in the cat using the orthograde autoradiographic tracing method and the retrograde horseradish peroxidase technique. This was done in the following manner. First, the division of the facial nucleus that innervates the pinna muscles was determined by injecting the pinna muscles with HRP and surveying the facial nucleus for retrogradely filled cells. Second, the brainstem regions that project the facial nucleus were identified using the horseradish peroxidase method. Third, the superior colliculus projections to these areas were studied using the autoradiographic tracing method. The results suggest that superior colliculus control of pinna movements is mediated entirely by indirect connections with the facial nucleus and that these connections occur mainly in a paralemniscal zone in the lateral midbrain. Of all the brainstem regions shown by the horseradish peroxidase experiments to project to the facial nucleus only this midbrain paralemniscal zone received a projection from the superior colliculus that was dense and overlapped precisely the region containing facial projecting neurons. Further autoradiographic tracing revealed that the facial nucleus was the primary brainstem target of this paralemniscal zone and that all paralemniscal fibers projecting to the facial nucleus ended in the subdivision that innervates the pinna muscles. Other paralemniscal efferents terminate in the opposite paralemniscal zone. The data suggest that other connections between the superior colliculus and the facial nucleus may occur in the cuneiform nucleus of the midbrain, the region around the oculomotor complex, and the reticular formation dorsal to the superior olive.

Banding of lateral superior olivary nucleus afferents in the inferior colliculus: a possible substrate for sensory integration.

In this study the organization of the projection from the lateral superior olivary nucleus (LSO) to the inferior colliculus was investigated in the cat by using anterograde tract-tracing techniques. The findings indicated that LSO projected bilaterally to the central nucleus of the inferior colliculus as well as to the ventrolateral and rostral pole nuclei. In the central nucleus a larger medial component of the projection ended in pars medialis and centralis. A smaller lateral component ended in the region of the pars lateralis. Both components of the projection appeared to be topographically organized, but in the lateral component the low-frequency part of LSO appeared to have greater representation. The uncrossed and crossed LSO projections to the inferior colliculus exhibited several important differences in their distribution. First, periodic bands of dense labeling were more prominent in the distribution of the uncrossed projection. The bands measured 150-200 micron in thickness and in some cases interruptions or gaps were present along the length of the bands. The distribution of the crossed projection was more diffuse, but some banding was also apparent. Second, the positions of the bands of dense labeling on the two sides were not homotopic as determined by labeling projections from the ipsilateral and contralateral LSO in the same tissue. The dense bands labeled with WGA-HRP from an injection in LSO on one side and bands labeled with 3H-leucine from an injection in LSO on the other side either were interdigitating or were only partially overlapping. Finally, the area over which the uncrossed projection distributed endings varied in size with respect to that of the crossed projection. The variation in size of the area of the projections was a function of the frequency representation. A model based on the three-dimensional reconstruction of bands as projection sheets is proposed as a substrate for selective integration of afferents in the inferior colliculus.

Superior colliculus connections with the extraocular motor nuclei in the cat.

Direct and indirect projections from the cat superior colliculus to the extraocular motor nuclei were studied using the orthograde autoradiographic tracing method, the retrograde horseradish peroxidase technique, and Golgi methods. The results show that the superior colliculus projects to the central gray matter directly overlying the oculomotor complex. This projection arises almost entirely from the rostral third of the colliculus, and it terminates most heavily over the rostral half of the oculomotor complex. Dendrites of oculomotor cells extend into this tectal termination zone, making direct tecto-oculomotor contacts possible. Central gray cells within this termination zone project bilaterally to the abducens nuclei. It is proposed that the superior colliculus projection to the supraoculomotor central gray matter and the projection from the central gray matter to the abducens nuclei play a role in convergent eye movements. The superior colliculus projects lightly to a cell group directly ventrolateral to the trochlear nucleus. The superior colliculus sends a small direct projection to the contralateral abducens nucleus and a substantial projection to wide regions of the reticular formation that have been shown previously to project, in turn, to the abducens nucleus. Colliculus cells projecting to the abducens nucleus and adjacent reticular formation are located only in the caudal three-fourths of the colliculus, where they become increasingly concentrated at successively more caudal levels. It is proposed that the graded density of the cells of origin of this projection is the basic structural mechanism by which the colliculus generates horizontal foveating saccades of different amplitudes. Laminar analysis of the origin of all the superior colliculus projections to the extraocular motor regions described here revealed that they arise mostly from the stratum griseum intermedium.

Organization of the efferent projections of the medial superior olivary nucleus in the cat as revealed by HRP and autoradiographic tracing methods.

Features of the organization of the efferent axonal projections from the medial superior olivary nucleus (MSO) in the cat were studied. In order to determine the origin and distribution of projections from MSO, the retrograde horseradish peroxidase (HRP) and autoradiographic tracing methods were used. The results showed that (1) in both HRP and autoradiographic studies the projection to the inferior colliculus was largely ipsilateral, although a contralateral component was present; (2) the projection field of MSO was confined to the ventral division of the central nucleus of the inferior colliculus, and within this field the labeling was heavier in the rostral and dorsolateral parts of the ventral division; (3) the projection to the inferior colliculus was topographic with ventral parts of MSO projecting ventrally and dorsal parts of MSO projecting dorsolaterally; (4) the projection field in the central nucleus formed successive laminae oriented from ventrolateral to dorsomedial; (5) the axonal course was via the medial or internal segment of the lateral lemniscus; and (6) some fibers in this course ended additionally within the dorsal nucleus of the lateral lemniscus. This latter projection was also topographically organized. These observations supported previously described features of lamination and tonotopic order for afferents of the inferior colliculus, as well as recent suggestions that functional segregation of afferent connections exists within the laminated portion of the central nucleus of the inferior colliculus.

Nucleus sagulum: projections of a lateral tegmental area to the inferior colliculus in the cat.

The nucleus sagulum, an area of the midbrain tegmentum, has been considered a component of a lateral tegmental system within the ascending auditory pathway to the thalamus. In this study, connections of the nucleus sagulum within the midbrain were investigated in adult cats. Tracing methods using anterograde and retrograde axonal transport of markers were employed. The nucleus sagulum was identified as a region of principally small neurons (261 +/- 79 micron2) at the margin of the midbrain and neighboring the nuclei of the lateral lemniscus. Injections of tritiated leucine in the nucleus sagulum labeled axons that ended in dense patches within the superficial layers of the caudal portion of the dorsal cortex of the inferior colliculus on the ipsilateral side. Retrograde experiments confirmed this connection. Other axonal projections labeled in the anterograde studies included fibers ending in the dorsomedial nucleus, the superficial layers of the dorsal cortex, and the rostral nucleus of the inferior colliculus with some bilateral distribution. Outside of the inferior colliculus, sagulum injections labeled other axons ending in the ventral intercollicular tegmentum on both sides and in a dorsal and rostral region of the contralateral nucleus sagulum that appeared contiguous with the dorsal nucleus of the lateral lemniscus. The latter region included a population of larger neurons (340-540 micron2) and had different connections with the inferior colliculus. The distribution of axonal labeling after injections in the nucleus sagulum was contrasted with the distribution of projections from several neighboring areas of the lateral tegmentum, including the dorsal nucleus of the lateral lemniscus. None of these areas exhibited connections with the superficial layers of the caudal cortex of the inferior colliculus, which was the major target in the inferior colliculus of the nucleus sagulum. Thus, the results indicated that the nucleus sagulum is distinguished from adjacent regions of the lateral tegmentum by its connectivity. Its association with midbrain auditory pathways is supported by these connections as well as ascending ones to the auditory thalamus.

The vestibular complex of the American opossum didelphis virginiana. II. Afferent and efferent connections.

We have demonstrated the connectivity of the opossum's vestibular nuclei using degeneration, autoradiographic and horseradish peroxidase techniques and have found it to be generally comparable to that reported for the cat. Apart from the primary input described in Part I of our study, the cerebellum provides the major source of afferent connections to the vestibular complex. Axons from the cerebellar cortex distribute mainly to vestibular areas which receive no primary afferent projections, e.g., the dorsal part of the lateral vestibular nucleus, the dorsolateral margin of the inferior vestibular nucleus as well as cell groups comparable to "f" and "x." In contrast, fastigial fibers show considerable overlap with primary vestibular input, particularly in the ventral part of the lateral nucleus, the central part of the inferior nucleus and the medial nucleus. Axons of fastigial origin also distribute to the superior vestibular nucleus, to subnuclei "f" and "x" and to the parasolitary region. Although spinal fibers are diffuse within the main vestibular nuclei, they ramify quite densely within subnucleus "x." Most of the spinovestibular projection appears to arise in the cervical spinal cord of the opossum. Ipsilateral connections from the interstitial nucleus of Cajal and surrounding areas end predominantly, but not exclusively, in the medial vestibular nucleus. A crossed midbrain projection has been verified from the red nucleus to cell group "x" and the lateral part of the inferior nucleus, as well as to an area possibly comparable to cell group "z," as described for the cat. In Part I of our study we have shown that the major targets of primary vestibular fibers are the central part of the superior nucleus, a portion of the parabrachial complex possibly comparable to subnucleus "y"," the ventral part of the lateral nucleus and the medial nucleus. All of these primary zones give rise to fibers supplying extraocular nuclei and surrounding areas (present study). The ascending midbrain fibers from the superior nucleus end mainly ipsilaterally, whereas those from the putative subnucleus "y" and the medial vestibular nucleus distribute contralaterally for the most part. Although the dorsal part of the lateral vestibular nucleus has no primary vestibular input, it does receive a major projection from the cerebellar cortex. This same region of the lateral nucleus projects to the spinal cord, but not to extraocular nuclei. The ventral part of the lateral nucleus, and perhaps the medial nucleus, also relay to the spinal cord. Additional projections from all vestibular nuclei to the reticular formation provide indirect routes through which the vestibular nuclei can potentially effect multiple systems, including those innervating the spinal cord. Finally, commissural vestibular connections of the opossum are shown to arise within all four major nuclei.

Topographical organization of the inferior collicular projection and other connections of the ventral nucleus of the lateral lemniscus in the cat.

The topographic distribution of projections from the ventral nucleus of the lateral lemniscus (VNLL) in the cat was investigated with the autoradiographic tracing method. The origin of minor projections was verified by retrograde tracing methods. Small injections of tritiated leucine were placed in restricted zones of VNLL. A major afferent fiber system to the inferior colliculus was labeled in all cases. From the injection site labeled fibers coursed through and around the nuclei of the lateral lemniscus to enter the ipsilateral inferior colliculus. Regardless of the position or small size of the injection, labeled fibers distributed widely in the inferior colliculus. Fibers ended in the central nucleus and deeper layers of the dorsal cortex in most cases. There was also labeling in the ventrolateral nucleus, but very few fibers ended as lateral as the lateral nucleus. A small number of labeled fibers passed from the inferior colliculus into the nucleus of the brachium of the inferior colliculus and adjacent tegmental areas. Some labeled fibers entered the commissure of the inferior colliculus where they were traced into the dorsal cortex and rostral pole of the inferior colliculus on the side contralateral to the injection site. Though the projections labeled in individual cases were similar in their divergent pattern within the central nucleus of the inferior colliculus, specific variations in the pattern were found. The dorsal zone of VNLL projected more heavily to the deeper layers of the dorsal cortex and an adjacent field in the central nucleus than the other zones. Dorsal injections in the middle zone of VNLL, on the other hand, labeled the medial part of the central nucleus more heavily, whereas ventral injections in the middle zone resulted in heavier lateral labeling. The ventral zone of VNLL projected heavily to a central field in the central nucleus. In addition to this major afferent system of VNLL to the inferior colliculus, a smaller descending projection was found. The descending projection ended mainly in the dorsomedial periolivary region and ventral nucleus of the trapezoid body. However, in some cases a few fibers were traced to the cochlear nuclei. Finally, we observed projections to the medial geniculate body from the dorsal and ventral zones of VNLL that ended diffusely in the medial division of the medial geniculate body. Possibly some fibers from the dorsal zone contribute to a broader projection of the lateral tegmentum to the dorsal division of the medial geniculate body.

Development of glycinergic cells and puncta in nuclei of the superior olivary complex of the postnatal ferret.

The distribution of glycine-immunopositive cells and axonal endings was studied in the adult and early postnatal ferret superior olive. As in other species, the most prominent glycine-immunopositive cell group in the adult ferret superior olive was the medial nucleus of the trapezoid body. Other darkly immunostained cells were present, although more scattered, in most periolivary regions, including the lateral and ventral trapezoid body nuclei. In the lateral superior olivary nuclei, glycine-immunopositive cells were intermingled with immunonegative cells. A comparable population of cells in the ipsilateral lateral superior olivary nucleus was retrogradely labeled in cases with unilateral injections of tritiated glycine in the inferior colliculus. Glycine-immunopositive puncta were widely distributed in the neuropil in most periolivary regions, including dense accumulations in the dorsomedial periolivary region and ventral and lateral nuclei of the trapezoid body. In the lateral and medial superior olivary nuclei, immunopositive puncta were distributed around the principal cells in characteristic perisomatic halos. In postnatal ferrets, immunopositive cell bodies were first observed by postnatal day 7 and were distributed in regions comparable to regions in the adult, with the exception that immunopositive cells in the lateral superior olivary nucleus did not appear until about postnatal day 28. There was diffuse staining in the neuropil in principal and periolivary nuclei by postnatal day 7. During the third postnatal week, the immunostaining in the neuropil began to take on a more granular appearance and immunopositive puncta could be seen by postnatal day 35. In the lateral and medial superior olivary nuclei, the earliest distribution of immunostaining in the neuropil was nonuniform, being greater in the high-frequency, medial, and ventral regions, respectively. The density gradient in these areas was gradually eliminated over the next 2 postnatal weeks as immunostained processes and endings appeared over greater portions of the nuclei.

Laterality of superior olive projections to the inferior colliculus in adult and developing ferret.

The laterality of projections from the lateral superior olivary nucleus (LSO) to the inferior colliculus was studied in adult and immature postnatal ferrets. In the adult ferret, large unilateral injections of horseradish peroxidase (HRP) in the inferior colliculus labeled about equal proportions of cells in the ipsilateral and contralateral lateral superior olivary nuclei. The contralateral labeled cells consistently were more densely labeled than those on the ipsilateral side. Double labeling experiments using fluorescent dyes indicated that only about 3% of LSO cells in the adult give rise to collaterals ending in the inferior colliculus on both sides. As expected, the distribution of labeled cells varied topographically in the LSO as a function of the injection site in the inferior colliculus. Dorsolateral inferior collicular injections labeled cells in the lateral limb of the LSO, whereas ventromedial injections labeled cells in the medial limb of the LSO. The proportion of ipsilateral and contralateral labeled cells also varied across the lateral-medial axis of the LSO in some cases. A gradient in laterality was observed in these cases with the lateral limb of the LSO containing the highest proportion of contralateral labeled cells, and the medial limb, the highest proportion of ipsilateral labeled cells. Larger inferior collicular injections resulted in greater proportions of ipsilateral labeling in LSO than smaller injections. Finally, ipsilateral labeled cells tended to be in the marginal region of the LSO, whereas contralateral labeled cells were more common within the core region of the LSO, irrespective of the location along the lateral-medial axis of LSO. The contralateral predominance of labeled cells, greater density of labeling in contralateral cells, different topographic distribution, and regional segregation of ipsilateral and contralateral labeled cells were typical of the LSO in ferret kits by birth, one month before the onset of hearing. Nevertheless, the relative proportion of ipsilateral and contralateral projection cells appears to change during postnatal development.

Dendritic morphology and development in the ferret lateral superior olivary nucleus.

The dendritic morphology of cells in the lateral superior olivary nucleus was studied with the Golgi method in adult and postnatal ferrets. The lateral superior olivary nucleus in the adult ferret is a convoluted structure with an M-shape in frontal sections. The major cell type appears to have disk-shaped dendritic trees. Most dendritic trees appear to be approximately orthogonal to the curved medial-lateral axis of the nucleus. Depending on their position in the limb and on the plane of section with respect to the dendritic tree, the disk-shaped cells are either bipolar or radiate in orientation. One subclass of disk-shaped cells has secondary dendritic branches that end as tufts of tendril-like processes. In a second subclass of cells, the dendrites exhibit several orders of dichotomous branching and lack obvious tufts of terminal processes. Marginal cells are observed at the border of the nucleus and have dendrites restricted to the margins of the cell plate. The bipolar orientation of disk-shaped cells orthogonal to the axis of the limbs is already apparent by the time of birth. Transient spines and other appendages are abundant on somata and dendrites during the first postnatal week. By the end of the first postnatal month only distal appendages are found. Tufts of fine tendril-like processes appear at the ends of dendrites between postnatal days 28 and 56.

Connections of the dorsal nucleus of the lateral lemniscus: an inhibitory parallel pathway in the ascending auditory system?

This study examines the dorsal nucleus of the lateral lemniscus (DNLL) and its afferent and efferent connections. In Nissl-stained material, DNLL has three parts: dorsal, ventral, and lateral. Although each part contains neurons with similar Nissl patterns, the subdivisions may be distinguished by the size, shape, and orientation of the cells. The lateral DNLL contains a mixture of DNLL neurons and cells from the sagulum. Afferent connections to DNLL were investigated with anterograde axonal transport techniques. Bilateral inputs to DNLL arise from the anteroventral cochlear nucleus and lateral superior olive, while unilateral inputs are provided by the ipsilateral medial superior olive and the contralateral DNLL. The inputs appear to have a tonotopic organization. Afferent fibers to DNLL form horizontal bands that are continuous both mediolaterally and rostrocaudally. All parts of DNLL do not share the same inputs, and a medial-to-lateral gradient in the labeling of some pathways is evident. To study the efferent connections of DNLL, both retrograde and anterograde axonal transport techniques were used. The DNLL projects to the inferior colliculus and the contralateral DNLL. The topography of these projections suggests that areas of similar tonotopic organization are connected. In the inferior colliculus, the projection is heaviest to the central nucleus and extends to the adjacent dorsal and caudal cortex, the rostral pole nucleus, and the ventrolateral nucleus. Axons from DNLL terminate along the fibrodendritic laminae of the central nucleus as bands that are prominent on the contralateral side, whereas those on the ipsilateral colliculus are more diffuse. The afferent and efferent connections of DNLL constitute a multisynaptic pathway, parallel to the other ascending pathways to the inferior colliculus. The other ascending pathways include the direct pathways from the cochlear nucleus to the inferior colliculus and the indirect pathways via the superior olivary complex. Ascending pathways are discussed as to their relationship to the subdivisions of the inferior colliculus, the laterality of their projections, and their banding patterns in the central nucleus. In contrast to the excitatory pathways to the inferior colliculus, the neurons in DNLL may use GABA as a neurotransmitter. Axons from the DNLL terminate in the inferior colliculus as bands that could have a unique inhibitory function. Thus, the multisynaptic, DNLL pathway may provide feed-forward inhibitory inputs to the inferior colliculus, bilaterally, and to the contralateral DNLL.