Descending projections from the inferior colliculus to medial olivocochlear efferents: Mice with normal hearing, early onset hearing loss, and congenital deafness.

Auditory efferent neurons reside in the brain and innervate the sensory hair cells of the cochlea to modulate incoming acoustic signals. Two groups of efferents have been described in mouse and this report will focus on the medial olivocochlear (MOC) system. Electrophysiological data suggest the MOC efferents function in selective listening by differentially attenuating auditory nerve fiber activity in quiet and noisy conditions. Because speech understanding in noise is impaired in age-related hearing loss, we asked whether pathologic changes in input to MOC neurons from higher centers could be involved. The present study investigated the anatomical nature of descending projections from the inferior colliculus (IC) to MOCs in 3-month old mice with normal hearing, and in 6-month old mice with normal hearing (CBA/CaH), early onset progressive hearing loss (DBA/2), and congenital deafness (homozygous Shaker-2). Anterograde tracers were injected into the IC and retrograde tracers into the cochlea. Electron microscopic analysis of double-labelled tissue confirmed direct synaptic contact from the IC onto MOCs in all cohorts. These labelled terminals are indicative of excitatory neurotransmission because they contain round synaptic vesicles, exhibit asymmetric membrane specializations, and are co-labelled with antibodies against VGlut2, a glutamate transporter. 3D reconstructions of the terminal fields indicate that in normal hearing mice, descending projections from the IC are arranged tonotopically with low frequencies projecting laterally and progressively higher frequencies projecting more medially. Along the mediolateral axis, the projections of DBA/2 mice with acquired high frequency hearing loss were shifted medially towards expected higher frequency projecting regions. Shaker-2 mice with congenital deafness had a much broader spatial projection, revealing abnormalities in the topography of connections. These data suggest that loss in precision of IC directed MOC activation could contribute to impaired signal detection in noise.

Effects of long-term salicylate administration on synaptic ultrastructure and metabolic activity in the rat CNS.

Tinnitus is associated with neural hyperactivity in the central nervous system (CNS). Salicylate is a well-known ototoxic drug, and we induced tinnitus in rats using a model of long-term salicylate administration. The gap pre-pulse inhibition of acoustic startle test was used to infer tinnitus perception, and only rats in the chronic salicylate-treatment (14 days) group showed evidence of experiencing tinnitus. After small animal positron emission tomography scans were performed, we found that the metabolic activity of the inferior colliculus (IC), the auditory cortex (AC), and the hippocampus (HP) were significantly higher in the chronic treatment group compared with saline group (treated for 14 days), which was further supported by ultrastructural changes at the synapses. The alterations all returned to baseline 14 days after the cessation of salicylate-treatment (wash-out group), indicating that these changes were reversible. These findings indicate that long-term salicylate administration induces tinnitus, enhanced neural activity and synaptic ultrastructural changes in the IC, AC, and HP of rats due to neuroplasticity. Thus, an increased metabolic rate and synaptic transmission in specific areas of the CNS may contribute to the development of tinnitus.

Tectothalamic inhibitory projection neurons in the avian torus semicircularis.

Inhibitory feedforward projection is one of key features of the organization of the central auditory system. In mammals, the inferior colliculus (IC) is the origin of a substantial inhibitory feedforward projection as well as an excitatory projection to the auditory thalamus. This inhibitory feedforward projection is provided by large γ-aminobutyric acid (GABA)ergic (LG) neurons, which are characterized by their receipt of dense excitatory axosomatic terminals positive for vesicular glutamate transporter (VGLUT) 2. In the avian torus semicircularis (TS), which is the homolog of the IC, neither the homology of cell types nor the presence of inhibitory feedforward inhibition have been established. In this study, we tested the presence of LG neurons in pigeon and chicken by neuroanatomical techniques. The TS contained two types of GABAergic neurons of different soma size. Of these, larger GABA + cells were encircled by dense VGLUT2 + axosomatic terminals. Ultrastructural analyses revealed that more than 30% of the perimeter of a large GABA+, but not small GABA + or GABA-, soma was covered by presumptive excitatory axosomatic terminals, suggesting that large GABA + cells are the sole recipient of dense excitatory axosomatic synapses. After injection of a retrograde tracer into the auditory thalamus, many retrogradely labeled neurons were found bilaterally in the TS, a few of which were GABA+. Almost all tectothalamic GABA + neurons had large somata, and received dense VGLUT2 + axosomatic terminals. These results clearly demonstrated the presence of LG neurons in birds. The similar morphology of LG neurons implies that the function of tectothalamic inhibition is similar among amniotes. J. Comp. Neurol. 524:2604-2622, 2016. © 2016 Wiley Periodicals, Inc.

Analysis of excitatory synapses in the guinea pig inferior colliculus: a study using electron microscopy and GABA immunocytochemistry.

The inferior colliculus (IC) integrates ascending auditory input from the lower brainstem and descending input from the auditory cortex. Understanding how IC cells integrate these inputs requires identification of their synaptic arrangements. We describe excitatory synapses in the dorsal cortex, central nucleus, and lateral cortex of the IC (ICd, ICc and IClc) in guinea pigs. We used electron microscopy (EM) and post-embedding anti-GABA immunogold histochemistry on aldehyde-fixed tissue from pigmented adult guinea pigs. Excitatory synapses were identified by round vesicles, asymmetric synaptic junctions, and gamma-aminobutyric acid-immunonegative (GABA-negative) presynaptic boutons. Excitatory synapses constitute ∼60% of the synapses in each IC subdivision. Three types can be distinguished by presynaptic profile area and number of mitochondrial profiles. Large excitatory (LE) boutons are more than 2 µm(2) in area and usually contain five or more mitochondrial profiles. Small excitatory (SE) boutons are usually less than 0.7 µm(2) in area and usually contain 0 or 1 mitochondria. Medium excitatory (ME) boutons are intermediate in size and usually contain 2 to 4 mitochondria. LE boutons are mostly confined to the ICc, while the other two types are present throughout the IC. Dendritic spines are the most common target of excitatory boutons in the IC dorsal cortex, whereas dendritic shafts are the most common target in other IC subdivisions. Finally, each bouton type terminates on both gamma-aminobutyric acid-immunopositive (GABA+) and GABA-negative (i.e., glutamatergic) targets, with terminations on GABA-negative profiles being much more frequent. The ultrastructural differences between the three types of boutons presumably reflect different origins and may indicate differences in postsynaptic effect. Despite such differences in origins, each of the bouton types contact both GABAergic and non-GABAergic IC cells, and could be expected to activate both excitatory and inhibitory IC circuits.

Long-term regulation in calretinin staining in the rat inferior colliculus after unilateral auditory cortical ablation.

In this study we analyzed the effects in the inferior colliculus of a unilateral ablation of the auditory cortex in rats. Variations in both calretinin immunoreactivity and protein levels determined by Western blot suggest that such lesions induce changes in the regulation of this calcium-binding protein. Stereological counts of calretinin-immunoreactive neurons in the inferior colliculus 15, 90, and 180 days after the lesion showed a progressive increase in the number of immunoreactive neurons, with a parallel increase in the intensity of staining. Two hundred forty days after the cortical lesion, both the number of immunoreactive neurons and the staining intensity had returned to control values. The effects of the cortical lesion on calretinin regulation are more intense in those inferior colliculus subdivisions more densely innervated by the corticocollicular projection. This finding, along with the time course of calretinin regulation suggests that degeneration of the descending projection is linked to calretinin regulation in the inferior colliculus. We hypothesize, based on the role of calretinin, that the observed increase in immunoreactivity levels seen in the inferior colliculus after lesioning of the auditory cortex may be related to altered excitability in deafferented neurons. Our finding, may reflect adaptive mechanisms to changes in calcium influx and excitability in inferior colliculus neurons induced by lesions of the descending projection from the cortex to the inferior colliculus.

Two classes of GABAergic neurons in the inferior colliculus.

The inferior colliculus (IC) is unique, having both glutamatergic and GABAergic projections ascending to the thalamus. Although subpopulations of GABAergic neurons in the IC have been proposed, criteria to distinguish them have been elusive and specific types have not been associated with specific neural circuits. Recently, the largest IC neurons were found to be recipients of somatic terminals containing vesicular glutamate transporter 2 (VGLUT2). Here, we show with electron microscopy that VGLUT2-positive (VGLUT2(+)) axonal terminals make axosomatic synapses on IC neurons. These terminals contain only VGLUT2 even though others in the IC have VGLUT1 or both VGLUT1 and 2. We demonstrate that there are two types of GABAergic neurons: larger neurons with VGLUT2(+) axosomatic endings and smaller neurons without such endings. Both types are present in all subdivisions of the IC, but larger GABAergic neurons with VGLUT2(+) axosomatic terminals are most prevalent in the central nucleus. The GABAergic tectothalamic neurons consist almost entirely of the larger cells surrounded by VGLUT2(+) axosomatic endings. Thus, two types of GABAergic neurons in the IC are defined by different synaptic organization and neuronal connections. Larger tectothalamic GABAergic neurons are covered with glutamatergic axosomatic synapses that could allow them to fire rapidly and overcome a slow membrane time constant; their axons may be the largest in the brachium of the IC. Thus, large GABAergic neurons could deliver IPSPs to the medial geniculate body before EPSPs from glutamatergic IC neurons firing simultaneously.

Learning drives differential clustering of axodendritic contacts in the barn owl auditory system.

Computational models predict that experience-driven clustering of coactive synapses is a mechanism for information storage. This prediction has remained untested, because it is difficult to approach through time-lapse analysis. Here, we exploit a unique feature of the barn owl auditory localization pathway that permits retrospective analysis of prelearned and postlearned circuitry: owls reared wearing prismatic spectacles develop an adaptive microcircuit that coexists with the native one but can be analyzed independently based on topographic location. To visualize the clustering of axodendritic contacts (potential synapses) within these zones, coactive axons were labeled by focal injection of fluorescent tracer and their target dendrites labeled with an antibody directed against CaMKII (calcium/calmodulin-dependent protein kinase type II, alpha subunit). Using high-resolution confocal imaging, we measured the distance from each contact to its nearest neighbor on the same branch of dendrite. We found that the distribution of intercontact distances for the adaptive zone was shifted dramatically toward smaller values compared with distributions for either the maladaptive zone of the same animals or the adaptive zone of normal juveniles, which indicates that a dynamic clustering of contacts had occurred. Moreover, clustering in the normal zone was greater in normal juveniles than in prism-adapted owls, indicative of declustering. These data demonstrate that clustering is bidirectionally adjustable and tuned by behaviorally relevant experience. The microanatomical configurations in all zones of both experimental groups matched the functional circuit strengths that were assessed by in vivo electrophysiological mapping. Thus, the observed changes in clustering are appropriately positioned to contribute to the adaptive strengthening and weakening of auditory-driven responses.

Synaptogenesis in the inferior colliculus of the pre-hearing postnatal ferret.

Although intrinsic organization in the inferior colliculus (IC) has been surveyed in a variety of species, current knowledge of synaptogenesis within the mammalian inferior colliculus is limited. The present study surveyed the ultrastructure of the central nucleus of the inferior colliculus in postnatal day (P) P4, P7, P14, and P28 ferrets, prior to the onset of hearing at the end of the first postnatal month with the goal of beginning to characterize the time course of synapse formation in relation to the development of afferent projection patterns within the IC. Results suggest that initial synaptogenesis has occurred in the IC by P4 and continues during the period when maturation of the distribution of axons from brainstem auditory nuclei is taking place.

Mossy fibers in granule cell areas of the rat dorsal cochlear nucleus from intrinsic and extrinsic origin innervate unipolar brush cell glomeruli.

Non tonotopic transmission between cochlear nuclei and other auditory and non-auditory nuclei in the brain is probably due to large axonal terminals (mossy fibers) innervating granule cell areas of cochlear nuclei. The origin of mossy fibers in the dorsal cochlear nucleus (DCN) is multiple, from other auditory or non-auditory nuclei but possibly also from intrinsic neurons. The present ultrastructural immunocytochemical study reports for the first time the presence of anterograde-labeled mossy fibers in the DCN of the rat after injection of the neural tracer WGA-HRP into 3 different nuclei. Labeled mossy fibers were seen in 9.0% of mossy fibers detected after tracer injection into the ipsilateral anteroventral cochlear nucleus, in 7.3% of mossy fibers after contralateral collicular injection, and 13.2% after contralateral cochlear nucleus injection. Most (over 95%) mossy fibers contained round vesicles, both large and small, and were likely excitatory terminals, but few showed flat-pleomorphic vesicles that contained the inhibitory neurotransmitters GABA and glycine. Most of the anterograde-labeled ipsilateral mossy fibers containing small round synaptic vesicles, are probably derived from multipolar neurons within the ipsilateral anteroventral cochlear nucleus. After injections into the contralateral inferior colliculus, it was not possible to distinguish putative descending collicular mossy fibers from intrinsic mossy fibers. The latter would suggest the presence of an amplification pathway within the DCN, from collateral axons of pyramidal or stellate cells of the ipsilateral ventral cochlear nucleus to form glomeruli with granule-unipolar brush cells. After injection into the contralateral cochlear nucleus, it was not possible to distinguish between commissural mossy fibers and those derived from ipsilateral recurrent axon-terminals of commissural neurons within the DCN or the ventral cochlear nucleus. Despite this limitation, the present observations show that extrinsic or intrinsic mossy fibers reach granule cell areas in layers 2 and 3 of the DCN and form glomeruli of large or small dimension (1.5-4 microm) with unipolar brush and granule cells. These mossy fibers probably carry a fast excitatory non-tonotopic input which may influence the electrical response of granule cell areas.

CNTFRalpha and CNTF expressions in the auditory brainstem: light and electron microscopy study.

CNTF receptor alpha (CNTFRalpha) is involved in the development, the maintenance and the regeneration of a variety of brain structures. However, its in vivo distribution has not been determined in the auditory system. CNTFRalpha expression was studied in developing and adult rat brainstem auditory nuclei using immunohistochemistry. At birth, the CNTFRalpha immunolabeling was clearly present in somata of the external nucleus of the inferior colliculus but was diffuse throughout brainstem auditory nuclei. The labeling was present in most brainstem auditory nuclei by post-natal day (PND) 6. The intensity of the staining subsequently increased to its highest level at PND21 and decreased to an adult-like appearance by the fourth post-natal week. In adult, CNTFRalpha labeling occurred in most neurons of the cochlear nucleus (CN), the lateral superior olive (LSO), the medial superior olive (MSO), and the medial nucleus of the trapezoid body (MNTB). CNTFRalpha labeling first appeared in the central nucleus of the inferior colliculus (IC) by the end of the fourth week. There was a general increase in the expression of CNTFRalpha that begins prior to the onset of hearing and reaches its highest level after this important developmental stage. Ultrastructural analysis in the adult ventral CN revealed the presence of CNTFR in post-synaptic sites. The presence of CNTF has been investigated in the adult using both Western blot and immunohistochemistry. Western blot showed the presence of CNTF in both peripheral and central auditory structures. The CNTF label was generally localized to the somatic compartment, in axons and as puncta surrounding neuronal cell bodies and dendrites. Differential CNTF labeling was observed between the different auditory nuclei. CNTF staining is present in neurons of the CN, the MNTB and the LSO, while it is restricted to axons and puncta surrounding neuronal somata in the IC. The clear presence of CNTFRalpha at post-synaptic terminals and that of its ligand the CNTF in axons and puncta surrounding neuronal cell bodies suggest an anterograde mode of action for CNTF in the central auditory system.

Parvalbumin-like immunostaining in the cat inferior colliculus. Light and electron microscopic investigation.

The presence of the calcium-binding protein parvalbumin (PV) was studied in neuronal elements of the cat's inferior colliculus (IC) by means of light and electron microscopic immunocytochemistry. Immunostaining of PV was detected in all three main parts of the IC. Several subtypes of large neurons that differed in size and shape were immunostained, comprising approx. 15% of the total number of PV-containing neurons. Approx. half of the labeled neurons were medium sized. Two types of small neurons were found to be PV synthesizing, and comprised approx. 35% of the total PV-containing population. Ultrastructurally, many dendrites were heavily immunolabeled, and the reaction product was present in dendritic spines as well. Several types of synaptic boutons contained reaction product, and terminated on both labeled and unlabeled postsynaptic targets forming asymmetric and symmetric synapses. Approx. 70% of all PV-immunolabeled terminals contained round synaptic vesicles and formed asymmetric synapses. The majority of these boutons were of the "large round" type and corresponded to the terminals of cochlear nuclei. A lower number were of the "small round" type, and were probably corticotectal terminals. The remaining 30% of PV-containing terminals contained pleomorphic or elongated vesicles and formed symmetric synapses. These terminals corresponded with "P" and "F1" bouton types. Part of these boutons appeared to arise from nuclei of the lateral lemniscus and the superior olive, and a certain percentage likely represented endings of inhibitory interneurons.

Ultrastructural immunocytochemistry for glycine in neurons of the dorsal cochlear nucleus of the guinea pig.

Neurons in the dorsal cochlear nucleus of the guinea pig were classified according to their positivity to the inhibitory neurotransmitter glycine, ultrastructure and projections to the inferior colliculus as indicated by tract-tracing and ultrastructural immunocytochemistry. Only some pyramidal and few giant cells, surrounded by glycinergic boutons containing flat and pleomorphic vesicles, projected to the inferior colliculus as glycine-negative excitatory cells. Smaller neurons in superficial layers of the dorsal cochlear nucleus did not project to the inferior colliculus, and were recognized as glycine-negative granule and unipolar brush cells. Few glycinergic, inhibitory neurons among granule cells were indicated as Golgi-stellate neurons. All small neurons associated to the granule cell areas received few, mainly glycinergic synapses, and their dendrites contacted large boutons (mossy fibers). Other medium-large glycine positive neurons in the superficial (cartwheel) and deep layers (tuberculo-ventral and large-giant) of the dorsal cochlear nucleus did not project to the inferior colliculus. Giant-large glycinergic neurons surrounded by sparse axo-somatic, mostly glycinergic synapses, probably represent commissural neurons projecting to the contralateral cochlear nucleus. Rare boutons, possibly descending from the inferior colliculus, were seen onto pyramidal cells or their dendrites, and these boutons mainly stored glycine positive pleomorphic vesicles or glycine negative round vesicles. No descending mossy fibers storing round vesicles were labelled from the central nucleus of the inferior colliculus. These observations suggest that very few terminals in the dorsal cochlear nucleus of the guinea pig are derived from the inferior colliculus.

Putative inhibitory collicular boutons contact large neurons and their dendrites in the dorsal cochlear nucleus of the rat.

Within the circuits of the acoustic nuclei, the inferior colliculus sends descending (collicular) terminals to control with a feedback mechanism, part of the activity of the dorsal cochlear nucleus (DCN). It is not known whether this descending projection is prevalently excitatory or inhibitory. Using the neuronal tracer Wheat Germ Agglutinin conjugated to Horse Radish Peroxidase (WGA-HRP) the connections between the inferior colliculus and the DCN of the rat have been investigated. By far most retrograde labelled large neurons were glycine and GABA negative (pyramidal and giant neurons) and rare medium-size cells were glycine positive. The ultrastructural immunocytochemical analysis for glycine and GABA shows that mainly large, excitatory, neurons innervate the inferior colliculus. Rare medium-size glycine-positive cells with intermediate characteristics between pyramidal and cartwheel cells, seem also to project to the colliculus. Few WGA-HRP labelled boutons contact the large cells or their dendrites, have symmetric pre- and post-synaptic thickenings, contain pleomorphic and/or flat vesicles, and are labelled for GABA or glycine. Since no GABA labelled cells in both the dorsal and ventral cochlear nucleus were retrograde labelled from the colliculus, the source of these intrinsic anterograde labelled boutons must be external to the cochlear nucleus. GABA positive neurons are both present in the inferior colliculus (injected with the tracer) and superior olivary complex (not injected with the tracer). This suggests that the double labelled boutons (WGA-HRP and GABA) are inhibitory GABA-ergic collicular terminals contacting the excitatory neurons of the DCN. Other few boutons or mossy fibers containing round vesicles and immunonegative for both glycine and GABA, were also seen contacting the large neurons and their dendrites in the DCN. As the round vesicles boutons may be derived from other retrograde cells of the cochlear nucleus (pyramidal and stellate cells) and those glycine positive from the glycinergic neurons in paraolivary nuclei, it is more likely that only the WGA-HRP and GABA labelled boutons are true collicular terminals.

Tyrosine hydroxylase-like immunoreactive synaptic boutons in the inferior colliculus of the cat.

The data on the distribution of catecholaminergic cells and fibers in such a significant subcortical relay auditory center as the inferior colliculus (IC) are both few and controversial, and ultrastructural data are lacking. Young adult mongrel cats of both sexes were used. Following routine preparation procedures, the ultrathin sections were prepared for the ultrastructural examination of tyrosine hydroxylase (TH)-like immunoreactivity. TH-positive neuronal perikarya were not detected in the IC. On the other hand, an appreciable number of TH-immunoreactive unmyelinated axons and synaptic boutons were found in all subdivisions of the IC, most often in the nucleus externus, followed by the nucleus pericentralis, and a few were seen in the dorsomedial part of the central nucleus. The boutons measured 0.5-1.8 microns, contained pleomorphic synaptic vesicles, and established symmetrical synaptic contacts almost exclusively with dendrites of small caliber.

Comparison of the fine structure of cortical and collicular terminals in the rat medial geniculate body.

Neurons throughout the rat medial geniculate body, including the dorsal and ventral divisions, display a variety of responses to auditory stimuli. To investigate possible structural determinants of this variability, measurements of axon terminal profile area and postsynaptic dendrite diameter were made on inferior colliculus and corticothalamic terminal profiles in the medial geniculate body identified by anterograde tracer labeling following injections into the inferior colliculus or cortex. Over 90% of the synapses observed were axodendritic, with few axosomatic synapses. Small (<0.5 microm(2)) and large (>1.0 microm(2)) collicular profiles were found throughout the medial geniculate, but were smaller on average in the dorsal division (0.49+/-0.49 microm(2)) than in the ventral division (0.70+/-0.64 microm(2)). Almost all corticothalamic profiles were small and ended on small-caliber dendrites (0.57+/-0.25 microm diameter) throughout the medial geniculate. A few very large (>2.0 microm(2)) corticothalamic profiles were found in the dorsal division and in the marginal zone of the medial geniculate. GABA immunostaining demonstrated the presence of GABAergic profiles arising from cells in the inferior colliculus. These profiles were compared with GABAergic profiles not labeled with anterograde tracer, which were presumed to be unlabeled inferior colliculus profiles or thalamic reticular nucleus profiles. The distributions of dendritic diameters postsynaptic to collicular, cortical and unlabeled GABAergic profiles were compared with dendritic diameters of intracellularly labeled medial geniculate neurons from rat brain slices. Our results demonstrate a corticothalamic projection to medial geniculate body that is similar to other sensory corticothalamic projections. However, the heterogeneous distributions of excitatory inferior collicular terminal sizes and postsynaptic dendritic diameters, along with the presence of a GABAergic inferior collicular projection to dendrites in the medial geniculate body, suggest a colliculogeniculate projection that is more complex than the ascending projections to other sensory thalamic nuclei. These findings may be useful in understanding some of the differences in the response characteristics of medial geniculate neurons in vivo.

Putative commissural and collicular axo-somatic terminals on neurons of the rat ventral cochlear nucleus.

The type of synaptic terminals from the cochlear nucleus and inferior colliculus that terminate in the contralateral ventral cochlear nucleus are not known. These terminals were studied with the electron microscope and immunogold after injection of wheat germ agglutinin conjugated to horseradish peroxidase into the inferior colliculus or into the cochlear nucleus. The tracer anterogradely labelled boutons onto the main neurons of the contralateral ventral cochlear nucleus. Most of these cells (95%) were glycine immuno-negative and represent excitatory neurons. After injection of the tracer into the contralateral inferior colliculus few anterogradely labelled boutons were seen on spherical and multipolar cells of type II in the anteroventral cochlear nucleus. Rare labelled boutons were present on multipolar cells of type I and II, globular neurons and octopus cells in the posteroventral cochlear nucleus. After injection into the contralateral dorsal and ventral cochlear nucleus labelled boutons were seen more frequently than after injection into the inferior colliculus. These terminals contacted most of large neurons, especially multipolar cells of type II and less frequently of type I. Also globular and spherical cells were contacted by commissural terminals. Octopus cells received less frequently putative commissural terminals. Most boutons contained pleomorphic vesicles and stored GABA. A lower number of boutons with pleomorphic and flat vesicles contained glycine and sometimes GABA, both inhibitory neurotransmitters. Few boutons containing round vesicles were immuno-negative for both glycine and GABA, and were considered putative commissural excitatory terminals. The latter often contacted glycinergic neurons of type II so that also these terminals might elicit an inhibition with at least a disynaptic mechanism after contralateral stimulation.

Cytology of large neurons in the guinea pig dorsal cochlear nucleus contacting the inferior colliculus.

Large neurons in the dorsal cochlear nucleus of the guinea pig which project to the inferior colliculus were identified after injections of the neural tracer WGA-HRP. Retrograde labelled cells (pyramidal and giant neurons) in the dorsal cochlear nucleus were glycine and GABA immunonegative and showed a similar ultrastructure. Between 30 and 60% of their perimeter was covered by axo-somatic boutons, most of which (>50%) contained pleomorphic synaptic vesicles. Other boutons (about 40% of total) contained flat vesicles and few (5-6%) contained round vesicles, a characteristic of the excitatory cells innervating the inferior colliculus. Immunogold-cytochemistry, coupled to silver intensification, showed that more than 50% of axo-somatic pleomorphic boutons and over 90% of boutons containing flat and pleomorphic vesicles store glycine. Rare WGA-HRP labelled axo-somatic boutons containing flat-pleomorphic vesicles were seen on pyramidal and giant neurons. This suggests that a few inhibitory collicular terminals contact the excitatory large neurons in the dorsal cochlear nucleus.

Age-related reductions in the activities of antioxidant enzymes in the rat inferior colliculus.

The inferior colliculus (IC) is a major relay and processing center of auditory signals in the midbrain and receives inputs from most other auditory nuclei. A number of studies have indicated age-related declines in the GABAergic and excitatory amino acid systems in the IC, including losses in both GABA immunoreactive (+) and GABA immunonegative (-) synapses. The goal of this project was to identify potential biochemical and morphological changes in the IC that may contribute to deficits in the functions of these neurotransmitters, using three age groups of Fischer-344 rats. Homogenates obtained from the IC showed age-dependent reductions in activities of the antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT), with a concomitant increase in lipid peroxidation. Dephosphorylation of IC homogenates with alkaline phosphatase reduced the activities of SOD and CAT in all age groups, which could be restored by protein kinase C (PKC)-dependent phosphorylation. Restoration of enzyme activity was specific to the PKC-alpha isozyme, but not to the beta1, beta2, delta or gamma forms. No age-dependent change in the levels of PKC isoforms (alpha, beta1, beta2 and gamma) was detectable in IC homogenates. Morphological analyses indicate decreases in mitochondrial density in the somata of both GABA+ and GABA- IC neurons in 19- and 28-month-old rats when compared to 3-month-olds, along with significantly higher matricial abnormalities. These data indicate age-related increases in oxidative stress in the IC, which could be partially restored by PKC. The progressive increase in oxidative stress with age may underlie changes in neuronal morphology and function of the IC.

Direct innervation of identified tectothalamic neurons in the inferior colliculus by axons from the cochlear nucleus.

The present study sought to identify tectothalamic neurons in the rat inferior colliculus that receive their innervation directly from the cochlear nuclei and to identify the axons that provide the innervation. A direct projection would bypass the binaural interactions of the superior olivary complex and provide the quickest route to the neocortex. Axons, primarily from the dorsal cochlear nucleus, were labeled with anterograde transport of dextran and terminated in the central nucleus of the inferior colliculus in a laminar pattern. Most labeled axons were thin and simply branched. Other axons were thicker, gnarly, less frequently observed and probably originated from the ventral cochlear nucleus. None had concentrated endbulbs or a nest of endings. Both types of axons terminated primarily in the central nucleus and layer 3 of the external cortex. This pattern suggests that the combination of these subdivisions in the rat are equivalent to the central nucleus as defined in other species. Tectothalamic neurons in the inferior colliculus in the same animals were identified by retrograde transport from the medial geniculate body and intracellular injection of Lucifer Yellow. A number of different cell types act as tectothalamic neurons and receive contacts from cochlear nucleus axons. These include flat cells (disc-shaped), less-flat cells and stellate cells. Two innervation patterns were seen: a combination of axosomatic and axodendritic contacts, and predominantly axodendritic contacts. Both patterns were seen in the central nucleus, but axosomatic contacts were seen less often in the other subdivisions. This is the first study to show direct connections between cochlear nuclear axons and identified tectothalamic neurons. The layers of axons from cochlear nuclei may provide convergent inputs to neurons in the inferior colliculus rather than the heavy inputs from single axons typical of lower auditory nuclei. Excitatory synapses made by axons from the cochlear nuclei on tectothalamic neurons may provide a substrate for rapid transmission of monaural information to the medial geniculate body.

Glycine-immunoreactive synapses in the central nucleus of the inferior colliculus. An electron microscopic study in the cat.

It has been known that the inferior colliculus contains many glycinergic fibers of both intrinsic and extrinsic nature. In the present study, glycine-immunoreactive (Gly-ir) synapses were examined in the central nucleus of the inferior colliculus in the cat. About half of 891 axondendritic synapses that were identified in the present study were Gly-ir. In 75% of these Gly-ir synapses, synaptic axon terminals contained pleomorphic or flattened synaptic vesicles and made symmetric synapses, while in 25% of Gly-ir synapses, synaptic axon terminals were filled with spherical synaptic vesicles and formed asymmetric synapses. Thus, Gly-immunoreactivity was detected in axodendritic synapses that formed both Gray's type I and Gray's type II synapses.