Contralateral Lateral Superior Olive Research Articles

Can auditory brainstem and midbrain processing of interaural level difference cues really explain perceptual performance?

Neurons in the lateral superior olive (LSO) are believed to be involved in processing of interaural level difference (ILD) cues for sound localization. However, the ILD-tuning of LSO neurons varies with the absolute sound pressure levels (SPLs) at the two ears, in contrast to the relatively robust perceptual processing of ILD cues at different absolute SPLs. Tsai et al. (J. Neurophysiol. 2010) proposed that if some inferior colliculus (IC) neurons compute the difference between the contralateral and ipsilateral LSO outputs, the dependencies on absolute SPLs could be canceled out. However, they only considered pure-tone stimulation of single neurons at their characteristic frequencies and at relatively low SPLs. In this study, we evaluated the hypothesis of Tsai and colleagues using computational models of populations of auditory nerve, LSO and IC neurons consistent with Tsai and colleagues’ proposal. Predictions from two different neural decoder algorithms applied to LSO and IC model outputs indicate that discharge-rate saturation and spread of excitation in the auditory nerve lead to even greater SPL-dependence of ILD coding across a population of LSO neurons than is apparent in single neurons, and the IC differencing operation proposed by Tsai and colleagues does relatively little to ameliorate this problem.

GluA4 sustains sensing of sounds through stable, speedy, sumptuous, spineless synapses

Glutamatergic synapses are of immense importance in that they mediate most excitation in the vertebrate brain. They are remarkably diverse. Some produce large, precisely timed synaptic currents, while others produce small, slow currents. Some are indefatigable, signalling robustly and with high fidelity, independent of how rapidly they are driven while others are plastic, constantly strengthening or weakening as a function of whether they are driven rapidly or slowly. α-Amino-3-hydroxy-5-methyl-4-isoxazole proprionate (AMPA) receptors report the presynaptic release of neurotransmitter to postsynaptic neurons. They are tetrameric ion channels formed as combinations of four α subunits, GluA1–4, each of which has the alternatively spliced forms, flip and flop. The subunits differ in the length and in the sequence of their cytoplasmic, carboxy terminal tails which mediate interactions with other molecular components of synapses that regulate receptor trafficking and tethering (Jackson & Nicoll, 2011). The receptor subunit composition determines the speed of synaptic signalling; its receptor number determines the strength of a synapse. It is essential for some functions of the brain that glutamatergic synapses not be altered by synaptic traffic. At the early stages of the auditory pathway, for example, acoustic information must be transmitted faithfully, without distortion by previous sounds, for that information to be useful for computing the location and meaning of sounds. Acoustic information from the cochlea is relayed through the cochlear nucleus and the contralateral medial nucleus of the trapezoid body (MNTB) to the contralateral lateral superior olive. The lateral superior olive compares the intensity of sounds at the two ears for localizing sound sources in the horizontal plane. MNTB neurons are excited through a single sumptuous, calyceal synapse that contacts the smooth surface of the target soma, the calyx of Held (Fig. 1A). This synapse has become a darling of synaptic physiologists because the presynaptic terminals are accessible to electrodes but has been largely ignored by cell biologists because it is boringly stable and not experimentally tractable. Figure 1 Glutamatergic, excitatory synapses are diverse As at upstream synapses, AMPA receptors in mature MNTB neurons are formed from GluA3 flop and/or GluA4 flop subunits in which GluA1 and GluA2 are not functionally evident (Joshi et al. 2004; Koike-Tani et al. 2005). In this issue of The Journal of Physiology, Yang et al. (2011) compare EPSCs in the MNTB of wild type mice with those in mice lacking GluA3 (GluA3 KO) and in mice lacking GluA4 (GluA4 KO). They find that in the MNTB of GluA4 KO mice, EPSCs are dramatically diminished in size and slowed whereas in GluA3 KO mice, EPSCs are only subtly slowed. GluA2 substituted to a small extent for the loss of GluA3 but not detectably for the loss of GluA4. The calyceal synapse does not rely entirely on GluA4, however, because the removal of GluA3 results in greater synaptic depression and desensitization than in the wild type. Subtle but detectable differences in presynaptic functions in KO mice serve as a reminder that differences between wild type and GluA4 KO mice reflect not only the removal of GluA4 at this synapse but can also reflect changes upstream in the auditory pathway as well as compensatory changes at all levels. These findings leave no doubt, however, that GluA4 is essential in mature, excitatory synapses in the MNTB, that GluA3 plays a minor role, and that GluA1 and 2 are almost undetectable in the MNTB. Cell biologists have been attracted by the dynamic glutamatergic synapses on spines in hippocampal pyramidal cells (Fig. 1B). They have studied the trafficking of AMPA receptors in hippocampal synapses where endogenous AMPA receptors are formed from either GluA1/2 or GluA2/3 subunits. Spines give neighbouring synapses biochemical privacy. The long, cytoplasmic, carboxy terminal tails of GluA1 subunits interact with proteins that drive AMPA receptors into synapses as a function of activity (Hayashi et al. 2000). GluA2/3 AMPA receptors cycle constitutively, replacing GluA1/2 receptors. The short cytoplasmic tails of GluA2 subunits regulate the interaction with a different group of synaptic proteins that promote the removal of GluA2-containing AMPA receptors (Kim et al. 2001). The relative rates of insertion and excision of receptors determines how many AMPA receptors are at a synapse and how much current it delivers at any moment. The findings of Yang et al. show that calyceal synapses of MNTB neurons manage without either GluA1 or GluA2, which are so important in the hippocampus, and do pretty well in the absence of GluA3. At these synapses fleets of the speedy GluA4 flop subunits seem to be coordinated by synaptic proteins in the communal postsynaptic environment of the soma.

A team of potassium channels tunes up auditory neurons

Mammals harbour a diverse array of voltage-gated K+ channels. How those channels team up in a neuron to do a job is becoming clear in the principal cells of the medial nucleus of the trapezoid body (MNTB). Ian Forsythe and his colleagues have used the electrically compact nature of MNTB neurons to measure K+ currents and combined those findings with molecular biological studies to identify, characterize and localize diverse K+ channels on MNTB neurons. A series of reports, the latest by Hardman and Forsythe being in this issue of The Journal of Physiology (Hardman & Forsythe, 2009), show that the firing of action potentials in MNTB neurons is finely controlled by a team of K+ conductances with different properties. Neurons in the MNTB are part of a neuronal circuit that assesses differences in intensity at the two ears, cues that are used to determine where in the horizontal plane sounds arise (Fig. 1A). Neurons in the lateral superior olive (LSO) compare interaural intensity by summing excitation from the ipsilateral ear with inhibition from the contralateral ear. Anyone who has participated in a conversation knows that sounds change rapidly in frequency and intensity; for interaural comparisons of intensity to be useful for localizing sounds, the time of arrival of excitation and inhibition at the LSO must be matched to within a few hundred microseconds. Figure 1 The medial nucleus of the trapezoid body (MNTB) feeds information about contralateral sounds to neurons in the lateral superior olive (LSO) that compare interaural intensity by balancing ipsilateral excitation (red) with contralateral inhibition (blue) ... How are the delays in ipsi- and contralateral inputs to the LSO matched in timing when path from the contralateral ear to the LSO is longer and includes an extra synapse? Neurons from the ipsilateral cochlear nucleus (CN) bring excitation to LSO neurons through relatively fine axons. Globular bushy cells from the contralateral CN, whose axons are particularly large and rapidly conducting, innervate MNTB neurons through calyces of Held. A calyx excites an MNTB neuron with a short and constant synaptic delay, generating large, suprathreshold EPSPs. The team of K+ channels allows MNTB neurons to relay the temporal firing pattern of globular bushy cells to contralateral LSO neurons as precisely timed, glycinergic IPSPs. Which are the members of the team of K+ channels? Voltage-gated K+ channels are formed a tetrameric combinations of α subunits from within a class of channels (e.g. Kv1). Figure 1B summarizes what is known about the presence of various α subunits in the MNTB. (1) Channels of the Kv1 family, located in the pre- and postsynaptic axons as well as on somas of MNTB neurons, provide a prominent, rapidly activating, low-voltage-activated K+ current whose activation around rest shapes synaptic potentials and prevents repetitive firing in both the calyceal terminals and in MNTB neurons (Dodson et al. 2003). (2) Channels that contain Kv2.2 are prominent in the initial segments of axons of MNTB neurons. These high-voltage-activated, slowly activating and deactivating channels hyperpolarize neurons between action potentials when they fire rapidly. In promoting repolarization between action potentials, these channels enhance Na+ currents by removing inactivation when MNTB neurons fire rapidly (Johnston et al. 2008b). (3) Channels containing Kv3.1b mediate a prominent high-voltage-activated, rapidly activating and inactivating K+ current that repolarizes action potentials, making them narrow. Kv3.1b channels are in calyceal terminals as well as on the somatic membranes of MNTB neurons. When they are phosphorylated, Kv3.1b subunits have a lower open probability. Rapid firing induced by electrical stimulation or by a noisy acoustic environment causes a decrease in phosphorylation and an increase in this K+ current (Song et al. 2005). (4) Kv4.3 subunits have been found in the MNTB of mice but not rats (Johnston et al. 2008a). Channels of the Kv4 family mediate A-currents. These K+ currents inactivate rapidly at relatively hyperpolarized potentials making them transient and most evident when depolarization is preceded by hyperpolarization. This small current would be expected to shorten and diminish EPSPs early in a train but becomes inactivated with strong stimulation. (5) Now Hardman and Forsythe show that Kv11 channels encoded by an ether-a-go-go-related gene (ERG) are also present in MNTB neurons of mice. These channels inactivate more rapidly than they activate so that they conduct current mainly upon repolarization after inactivation is removed but before they deactivate. The current through ERG channels repolarizes MNTB neurons, its action being most prominent near the resting potential after a depolarization. K+ channels work as a team to sharpen EPSPs, to sharpen action potentials, and to assure that repolarization removes Na+ inactivation even during rapid firing. K+ channels enable MNTB neurons to fire robustly and to signal with temporal precision even when they are driven at high rates.

Patterning of multiple layered projections to the auditory midbrain prior to experience

The precise arrangement of patterned inputs into discrete functional domains is a common organizational feature of primary sensory structures. While the specific organization of patterned connections has been well documented in the visual and somatosensory systems, comparatively little is known about the arrangement of neighboring afferent patterns in the emerging auditory system. Here we report early projection specificity for multiple converging inputs to the rat central nucleus of the inferior colliculus (ICC). Afferents arising from the dorsal cochlear nucleus (DCN), the dorsal nucleus of the lateral lemniscus (DNLL), and the lateral superior olive (LSO) establish discernible axonal layers a week prior to experience. By hearing onset, contralateral DCN and contralateral LSO layers are clearly defined and segregated from contralateral DNLL terminal zones. Layering of the ipsilateral LSO projection, on the other hand, exhibits considerable spatial overlap with the contralateral DNLL pattern. This fine laminar structure of interdigitating and overlapping inputs likely underlies the complex signal processing performed in the auditory midbrain and may serve as a model system for examining competitive interactions between neighboring excitatory and inhibitory projections early in development.

Anatomical projections of the nuclei of the lateral lemniscus in the albino rat (rattus norvegicus)

The ascending projections to the lateral lemniscal nuclei and the inferior colliculus were investigated in the albino rat by using Fluoro-Gold, either alone or in combination with other retrograde tract tracers. Injections were made into the central nucleus of the inferior colliculus (ICC), the dorsal nucleus of the lateral lemniscus (DNLL), the intermediate nucleus of the lateral lemniscus (INLL), or the ventral nucleus of the lateral lemniscus (VNLL). The ICC receives both ipsilateral and contralateral projections from the DNLL and the lateral superior olive, major ipsilateral projections from the INLL, VNLL, medial superior olive, and superior paraolivary nucleus, and major contralateral projections from both dorsal and ventral cochlear nucleus. The DNLL receives a similar pattern of projections from the auditory lower brainstem nuclei. The INLL, in contrast, receives its major projections from the ipsilateral VNLL, lateral superior olive, medial superior olive, superior paraolivary nucleus, and medial nucleus of the trapezoid body, but does not receive a heavy projection from the contralateral lateral superior olive. It receives a major contralateral projection from the ventral cochlear nucleus, but a much lighter projection from the contralateral dorsal cochlear nucleus. The VNLL receives projections from the ipsilateral medial nucleus of the trapezoid body and the contralateral ventral cochlear nucleus, but does not receive projections from the medial or lateral superior olives, the superior paraolivary nucleus, or the dorsal cochlear nucleus. Thus, the three primary subdivisions of the rat's lateral lemniscus can be distinguished from each other on the basis of their distinctive projection patterns.

Maturation of auditory brainstem projections and calyces in the congenitally deaf (dn/dn) mouse

The deaf dn/dn mouse is a valuable model of human congenital deafness. In this study we used the lipophylic dye DiA to trace auditory nerve and cochlear nucleus projections in the dn/dn mouse. In both normal and deaf mice, the ipsilateral projections from the anteroventral cochlear nucleus (AVCN) to the lateral superior olive (LSO), and the contralateral projections from the AVCN to the medial nucleus of the trapezoid body (MNTB) were intact. With age, there was a noted increase in the fenestration of the endbulb and calyx of Held, and this morphological maturation was also observed in the deaf mice, although there was a significant difference in total endbulb volume at P20 between normal and deaf mice. However, total calyceal volume was not significantly different between normal and deaf mice. There was electrophysiological evidence of in vivo spontaneous ventral cochlear nucleus activity in normal and deaf animals, indicating that this activity may be responsible for the appropriate connectivity in the deaf mice. Our results indicate that congenital deafness caused by the dn/dn mutation does not result in aberrant projections between the AVCN and the ipsilateral MNTB and contralateral LSO but can cause abnormalities in endbulb size.

Binaural Response Properties of Low-Frequency Neurons in the Gerbil Dorsal Nucleus of the Lateral Lemniscus

Differences in intensity and arrival time of sounds at the two ears, interaural intensity and time differences (IID, ITD), are the chief cues for sound localization. Both cues are initially processed in the superior olivary complex (SOC), which projects to the dorsal nucleus of the lateral lemniscus (DNLL) and the auditory midbrain. Here we present basic response properties of low-frequency (< 2 kHz) DNLL neurons and their binaural sensitivity to ITDs and IIDs in the anesthetized gerbil. We found many neurons showing binaural properties similar to those reported for SOC neurons. IID-properties were similar to that of the contralateral lateral superior olive (LSO). A majority of cells had an ITD sensitivity resembling that of either the ipsilateral medial superior olive (MSO) or the contralateral LSO. A smaller number of cells displayed intermediate types of ITD sensitivity. In neurons with MSO-like response ITDs that evoked maximal discharges were mostly outside of the range of ITDs the gerbil naturally experiences. The maxima of the first derivative of their ITD-functions (steepest slope), however, were well within the physiological range of ITDs. This finding is consistent with the concept of a population rather than a place code for ITDs. Moreover, we describe several other binaural properties as well as physiological and anatomical evidence for a small but significant input from the contralateral MSO. The large number of ITD-sensitive low-frequency neurons implicates a substantial role for the DNLL in ITD processing and promotes this nucleus as a suitable model for further studies on ITD-coding.

Molecular Plasticity in the Rat Auditory Brainstem: Modulation of Expression and Distribution of Phosphoserine, Phospho-CREB and TrkB after Noise Trauma

We induced acoustic trauma by applying click stimuli of 130 dB (SPL) for 30 min to one ear of adult rats. This treatment resulted in an instant and permanent threshold shift of 96 dB in the affected ear. A massive reduction of cochlear nerve fibers in the ventral cochlear nucleus (VCN) was demonstrated by tracing them from the cochlea of rats that survived acoustic overstimulation for 1 year or longer. In the auditory brainstem, we observed a deprivation-dependent appearance of fibers positive for tyrosine receptor kinase B in the ipsilateral VCN between day 3 and day 21 after trauma and an increase in phosphoserine immunostaining in the neuropil of the ipsilateral VCN and in neurons of the contralateral lateral superior olive during the first 30 days after trauma. Immunoreactivity for the cAMP response element binding protein in its phosphorylated form was transiently depressed in the ipsilateral inferior colliculus immediately after trauma and was elevated as late as 7 months after trauma in the ipsilateral VCN. Apparently, a unilateral acoustic overstimulation entails specific regulations of the activity of plasticity-associated molecules through phosphorylation and includes changes to neurotrophin signaling between neurons of the auditory brainstem.

Acoustic trauma induces reemergence of the growth- and plasticity-associated protein GAP-43 in the rat auditory brainstem.

We explored the consequences of unilateral acoustic trauma to intracochlear and central nervous system structures in rats. An acoustic trauma, induced by applying click stimuli of 130 dB (sound pressure level; SPL) for 30 minutes, resulted in an instant and permanent threshold shift of 95.92 +/- 1.08 dB (SEM) in the affected ear. We observed, as a consequence, a structural deterioration of the organ of Corti. Deprivation-dependent changes of neurons of the auditory brainstem were determined using antibodies against neurofilament and the growth-associated protein GAP-43 and compared with those following cochleotomy, studied earlier. By 231 days posttrauma, spiral ganglion cell bodies and their processes were almost entirely lost from all cochlear regions with destroyed organ of Corti. In the lateral superior olive (LSO) ipsilateral to the trauma, cell bodies of lateral olivocochlear neurons turned transiently GAP-43 positive within the first 1.5 years posttrauma. The time course of emergence and disappearance of this population of neurons was similar to that found after cochleotomy. Additionally, after noise trauma, principal cells in contralateral LSO and in medial superior olive (MSO) on both sides of the brainstem developed an expression of GAP-43 that began 3 and 16 days posttrauma, respectively, and lasted for at least 1 year. Such cells were rarely observed after cochleotomy. An unequivocal rise in GAP-43 immunoreactivity was also found in the neuropil of the inferior colliculus and the ventral cochlear nucleus, both preferentially on the acoustically damaged side. We conclude that the degree and specific cause of sudden unilateral deafness entail specific patterns of plasticity responses in the auditory brainstem, possibly to prevent the neural network dedicated to locate sounds in the environment from delivering erroneous signals centralward.

Ascending efferent projections of the superior olivary complex.

The superior olivary complex conveys information about binaural time and intensity to higher centers in the auditory pathway. This information is sent primarily to the subdivisions of the inferior colliculus and to the nuclei of the lateral lemniscus. Olivary projections are the predominant afferents to the central nucleus of the inferior colliculus. Electron microscopic observations of axonal endings in the central nucleus suggest that the ipsilateral medial superior olive and contralateral lateral superior olive make excitatory synapses. In contrast, the axons from the ipsilateral lateral superior olive to the central nucleus contain glycine and have a morphology consistent with inhibitory synapses. Little is known about the transmitter types used by olivary projections to the nuclei of the lateral lemniscus, but they are presumed to be similar to the collicular projections. Olivary ascending efferents are tonotopically organized and terminate in laminae in the inferior colliculus. They combine with other laminar afferents and postsynaptic neurons to create fibro-dendritic laminae in the colliculus. The key to the functional organization of the olivary efferents is the possible segregation of excitatory olivary efferents from each other in "synaptic domains" located on the laminae. This segregation may be the major determinant of response properties in the colliculus. Olivary efferents may converge with other non-olivary afferents on the same postsynaptic neurons in the colliculus. Inhibitory efferents from the lateral superior olive are essential in shaping the response properties of neurons in the colliculus. Olivary efferents to the nuclei of the lateral lemniscus are also key components of ascending pathways that inhibit neurons in the midbrain.

Glycine receptors in adult guinea pig brain stem auditory nuclei: regulation after unilateral cochlear ablation.

In young adult guinea pigs, the effects of unilateral cochlear ablation were determined on the specific binding of [3H]strychnine measured in subdivisions of the cochlear nucleus (CN), the superior olivary complex, and the auditory midbrain, after 2, 7, 31, 60, and 147 postlesion days. Changes in binding relative to that in age-matched controls were interpreted as altered activity and/or expression of synaptic glycine receptors. Postlesion binding declined ipsilaterally in most of the ventral CN and in the lateral superior olive (LSO). Binding was modestly deficient in the ipsilateral dorsal CN and in the anterior part of the contralateral anteroventral CN. Binding was elevated in the contralateral LSO. Transient changes also occurred. Binding was elevated transiently, between 2 and 31 days, contralaterally in parts of the anteroventral CN, bilaterally in the medial superior olive (MSO), and bilaterally in most of the midbrain nuclei. Binding was deficient transiently, at 60 days, in most of the contralateral CN and bilaterally in the midbrain nuclei. The present findings, together with previously reported postlesion changes in glycine release, were consistent with persistently weakened glycinergic inhibitory transmission ipsilaterally in the ventral CN and the LSO and bilaterally in the dorsal CN. Glycinergic inhibitory transmission was strengthened in the contralateral LSO and transiently strengthened in the MSO bilaterally. A hypothetical model of the findings suggested that glycine receptor regulation may depend on excitatory and glycinergic input to auditory neurons. The present changes in glycine receptor activity may contribute to altered auditory functions, which often accompany hearing loss.

Simultaneous anterograde labeling of axonal layers from lateral superior olive and dorsal cochlear nucleus in the inferior colliculus of cat

The laminar organization of the central nucleus of inferior colliculus includes layers of axons that may be important in shaping the responses of neurons. Depending on their source, some layered axons are afferents that are superimposed and terminate on the same postsynaptic neurons, while other layered afferents, such as those from the ipsilateral and contralateral lateral superior olive, terminate side-by-side. The specific pattern of convergence may dictate which populations of axons are presynaptic to layered disc-shaped neurons in the central nucleus. We compared the distribution of afferent axons from the dorsal cochlear nucleus and the lateral superior olive to the contralateral inferior colliculus in the cat. Injection sites in cochlear nucleus and superior olive were physiologically characterized by extracellular recordings of single and multiple units in response to monaural and binaural acoustic stimulation. Two separate injections were made in each case, and both injection sites contained units with overlapping best frequencies. Biotinylated dextran, fluorescent dextran, 3H-leucine, and wheat germ agglutinin conjugated to horseradish peroxidase were used as anterograde tracers. The present results show that layered axons from the dorsal cochlear nucleus and lateral superior olive are superimposed in part of the contralateral central nucleus. Both projections were arranged in rostro-caudally oriented axonal layers that converged in the ventral part of the central nucleus. However, in the dorsal part of the central nucleus, the same layer of axons from the dorsal cochlear nucleus did not terminate with afferents from the lateral superior olive. Within the overlapping layers in the ventral central nucleus, the overlap of axons from the dorsal cochlear nucleus and the lateral superior olive was uniform except for small patches that were usually smaller than the dendritic fields of disc-shaped neurons. These data suggest that the layers may create specific functional zones in the central nucleus of the inferior colliculus. One zone may contain neurons with binaural responses that combine the properties of the inputs from the contralateral lateral superior olive and the dorsal cochlear nucleus. A second zone may contain inputs from the cochlear nucleus but lack those of the lateral superior olive.

Linear and nonlinear encoding patterns for a two-tone stimulus in guinea pig auditory cortex

In young adult guinea pigs, the effects of unilateral cochlear ablation were determined on the specific binding of [3H]strychnine measured in subdivisions of the cochlear nucleus (CN), the superior olivary complex, and the auditory midbrain, after 2, 7, 31, 60, and 147 postlesion days. Changes in binding relative to that in age-matched controls were interpreted as altered activity and/or expression of synaptic glycine receptors. Postlesion binding declined ipsilaterally in most of the ventral CN and in the lateral superior olive (LSO). Binding was modestly deficient in the ipsilateral dorsal CN and in the anterior part of the contralateral anteroventral CN. Binding was elevated in the contralateral LSO. Transient changes also occurred. Binding was elevated transiently, between 2 and 31 days, contralaterally in parts of the anteroventral CN, bilaterally in the medial superior olive (MSO), and bilaterally in most of the midbrain nuclei. Binding was deficient transiently, at 60 days, in most of the contralateral CN and bilaterally in the midbrain nuclei. The present findings, together with previously reported postlesion changes in glycine release, were consistent with persistently weakened glycinergic inhibitory transmission ipsilaterally in the ventral CN and the LSO and bilaterally in the dorsal CN. Glycinergic inhibitory transmission was strengthened in the contralateral LSO and transiently strengthened in the MSO bilaterally. A hypothetical model of the findings suggested that glycine receptor regulation may depend on excitatory and glycinergic input to auditory neurons. The present changes in glycine receptor activity may contribute to altered auditory functions, which often accompany hearing loss.

Multiple projections from the ventral nucleus of the trapezoid body in the rat

An analysis of the central projections of the ventral nucleus of the trapezoid body (VNTB) in the rat, a region of the superior olivary complex known for its neuronal heterogeneity, was made using two anterograde axonal tracers, [ 3H]leucine and biotinylated dextran amine (BDA). A mixture of these tracers was injected iontophoretically into the VNTB and the results analyzed by first assessing magnitudes of autoradiographic signal in nuclei receiving projections and then identifying the axons and terminals responsible for this signal in parallel sets of sections processed for BDA. Our analysis showed that in addition to its projections to each cochlea via the olivocochlear bundle, the VNTB has 3 major central sites of axonal terminations: (1) the cochlear nucleus, particularly the molecular layer of the contralateral dorsal cochlear nucleus, (2) the contralateral lateral superior olive, and (3) the ipsilateral inferior colliculus. Other sites receiving projections from the VNTB included the VNTB itself and the nuclei of the lateral leminiscus. Significantly, the relative magnitudes of labeling within the nuclei receiving inputs from the VNTB varied consistently as a function of the dorsoventral location of the injection site, confirming previous work showing that there is a partial segregation within this nucleus of neurons according to their projections. Our data also revealed an orderly topographic pattern of projections to the cochlear nuclei, lateral superior olive and the inferior colliculus which is consistent with the known tonotopic organization both of the VNTB and these projection targets. Methodologically, the co-injection of two tracers was advantageous in that patterns of silver grains in autoradiographs could be used to confirm whether axons and terminals labeled with BDA had originated from labeled somata at the injection site or were the result of uptake of BDA by fibers of passage.

Axonal projections from the lateral and medial superior olive to the inferior colliculus of the cat: a study using electron microscopic autoradiography.

The superior olivary complex is the first site in the central auditory system where binaural interactions occur. The output of these nuclei is direct to the central nucleus of the inferior colliculus, where binaural inputs synapse with monaural afferents such as those from the cochlear nuclei. Despite the importance of the olivary pathways for binaural information processing, little is known about their synaptic organization in the colliculus. The present study investigates the structure of the projections from the lateral and medial superior olivary nuclei to the inferior colliculus at the electron microscopic level. Stereotaxic placement and electrophysiological responses to binaural sounds were used to locate the superior olive. Anterograde axonal transport of 3H-leucine was combined with light and electron microscopic autoradiography to reveal the location and morphology of the olivary axonal endings. The results show that the superior olivary complex contributes different patterns of synaptic input to the central nucleus of the inferior colliculus. Each projection from the superior olivary complex to the colliculus differs in the number and combinations of endings. Axonal endings from the ipsilateral medial superior olive were exclusively the round (R) type that contain round synaptic vesicles and make asymmetrical synaptic junctions. This morphology is usually associated with excitatory synapses and neurotransmitters such as glutamate. Endings from medial superior olive terminate densely in the central nucleus. The projection from the contralateral lateral superior olive also terminates primarily as R endings. This projection also includes small numbers of pleomorphic (PL) endings that contain pleomorphic synaptic vesicles and usually make symmetrical synaptic junctions. The PL morphology is associated with inhibitory synapses and transmitters such as gamma-aminobutyric acid and glycine. All endings from the contralateral lateral superior olive terminate much less densely than endings from the medial olive. In contrast, the projection from the ipsilateral lateral superior olive contributes both R and PL endings in roughly equal proportions. These ipsilateral afferents are heterogeneous in density and can terminate in lower or higher concentrations than endings from the contralateral side. These data show that the superior olive is a major contributor to the synaptic organization of the central nucleus of the inferior colliculus. The ipsilateral projections of the medial and lateral superior olive may produce higher concentrations of R endings than other inputs to the central nucleus. Such endings may participate in excitatory synapses. The highest concentrations of PL endings come from the ipsilateral lateral superior olive.(ABSTRACT TRUNCATED AT 400 WORDS)

Development of ventral cochlear nucleus projections to the superior olivary complex in gerbil

The postnatal development of the projection from the ventral cochlear nucleus to the principal nuclei in the superior olivary complex in gerbil (Meriones unguiculatus) was studied in an age-graded series of pups ranging from 0 to 18 days old. Small crystals of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) were inserted into the ventral cochlear nucleus of aldehyde-fixed brains, and the labeled projections were examined with epifluorescence microscopy. Selected sections were photooxidized in a solution of diaminobenzidine and subsequently processed for electron microscopy to examine the development of labeled synapses in the target nuclei. Horseradish peroxidase was injected into the ventral cochlear nucleus of adult gerbils to assess the form and persistence of projections observed in the neonatal animals. In addition, electrophysiological responses to acoustic stimuli of single units in the adult auditory brainstem were analyzed to confirm the functionality of the novel projection from the ventral cochlear nucleus to the contralateral lateral superior olive. By the day of birth (P0), developing axons from the ventral cochlear nucleus have already established highly ordered pathways to the three primary nuclei of the superior olivary complex: the ipsilateral lateral superior olive, the contralateral medial nucleus of the trapezoid body, and at the lateral and medial dendrites of the ipsilateral and contralateral medial superior olive, respectively. Developing axons from the ventral cochlear nucleus that innervated the contralateral medial nucleus of the trapezoid body lacked the terminal morphology characteristic of the calyx of Held, but began to adopt a more characteristic form on P5. The mature calyx appeared around P14-16. Exuberant developmental projections to topographically inappropriate areas of the superior olivary complex were not observed at the postnatal ages studied. In addition to the projections of the ventral cochlear nucleus to the superior olivary complex described in other species, we observed the development and maintenance of a major direct projection from the ventral cochlear nucleus to the contralateral lateral superior olive. On P0, ventral cochlear nucleus axons decussate in the dorsal trapezoid body, form a plexus at the dorsal edge of the contralateral medial superior olive, and enter the ventrolateral limb of the contralateral lateral superior olive. Over the next 2 weeks, fascicles of fibers form on the hilar and ventral aspects of the ventrolateral limb. Fibers arising from these fascicles form converging, but nonoverlapping, arborizations within the ventrolateral limb at right angles to the curvature of the nucleus. The medial region was devoid of labeled axons.(ABSTRACT TRUNCATED AT 400 WORDS)

Development of ectopic projections from the ventral cochlear nucleus to the superior olivary complex induced by neonatal ablation of the contralateral cochlea.

The ability of an animal to localize a sound in space requires the precise innervation of the superior olivary complex by the ventral cochlear nuclei on each side of the lower brainstem. This precise pattern of innervation could require an immutable recognition of appropriate targets by afferent processes arising from these nuclei. This possibility was investigated by destroying one cochlea of gerbil pups (Meriones unguiculatus) on the second postnatal day and assessing the projections from the ventral cochlear nucleus (VCN) on the unablated side to the superior olivary complex during the subsequent 2 weeks and after the animals had reached maturity. A crystal of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) was inserted into VCN on the unablated side in animals ranging in age from 3 to 14 days. To assess the permanence of any altered pattern of innervation, horseradish peroxidase was injected into VCN on the unablated side in adult, neonatally ablated animals. Finally, electrophysiological responses to acoustic stimuli delivered to the ear on the unablated side were recorded in the superior olivary complex of adult animals to assess whether altered innervation patterns were functional. Normative data were derived from our accompanying study of the development of VCN projections to the superior olivary complex in normal gerbils (Kil et al., this issue). Whereas VCN normally projects to the lateral aspect of the ipsilateral medial superior olive and to the medial aspect of the contralateral medial superior olive in control animals, in experimental animals VCN on the unablated side projects to both sides of these nuclei. Whereas in the gerbil, VCN normally projects only to the hilar area and to the ventrolateral limb of the contralateral lateral superior olive, in experimental animals VCN on the unablated side projects throughout this nucleus. This induced projection is specific in that the efferents to each limb of the contralateral nucleus are linked to the normal projection to the homotopic region of the ipsilateral nucleus. Whereas VCN innervates the contralateral medial nucleus of the trapezoid body in control animals, in experimental animals VCN on the unablated side provides calyces of Held in the ipsilateral nucleus as well. The induced projections to these three major subnuclei of the superior olivary complex first appear within 24 hours of the cochlear ablation and continue to develop over at least the subsequent 11 days. Thus, prior to the day when the cochlea becomes functional, VCN has established specific ectopic projections to loci normally innervated by VCN on the ablated side.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent reorganisation within the superior olivary complex of the gerbil: development and induction by neonatal, unilateral cochlear removal.

Cochlear removal in young animals has been shown to produce a variety of degenerative and generative effects within the auditory brainstem. A primary target for axons of neurons in the anteroventral cochlear nucleus (AVCN) is the superior olivary complex (SOC). Following unilateral cochlear removal in neonatal gerbils, AVCN neurons on the side of the removal die, and axons deriving from the AVCN on the unlesioned side produce new endings that innervate previously inappropriate target zones within the ipsilateral medial nucleus of the trapezoid body, both medial superior olives, and the contralateral lateral superior olive. In this study, we have used the anterograde transport of DiI and HRP from the AVCN to relate the formation of these endings to the time course of normal development in the gerbil brainstem. We have also examined the effects of cochlear removal at different ages, and survival to various ages after the removal, to define the time course for these generative phenomena. The results show that, while the major projection pathways from the AVCN to the SOC are in place at the time of birth, further and subtle development of AVCN terminal arbors occurs during the first postnatal week. This overlaps with the time during which cochlear removal produces the formation of exuberant afferents to the SOC from the intact AVCN. The exuberant afferents form through axon sprouting rather than through a suppression of normal, developmental regression. They appear to innervate tonotopically appropriate target regions within the SOC. The formation of the novel afferents begins within 3 days of the removal and appears to be complete within a further 5-7 days. By postnatal day (P) 10, both the normal development of the AVCN to SOC projection and the potential for alteration of that projection by removal of the contralateral cochlea appear to be over. These results suggest that the potential for forming novel projections in the gerbil auditory brainstem is lost before the onset of functional hearing (at P12) and is, therefore, unlikely to result from changes in auditory experience.

Evidence for glutamatergic projections from the cochlear nucleus to the superior olive and the ventral nucleus of the lateral lemniscus.

This study attempts to determine if projections ascending from the guinea pig cochlear nucleus (CN) could be glutamatergic and/or aspartatergic. Multiple radio frequency lesions were made to ablate the right CN. The ablation was verified histologically. To identify the principal targets of CN efferents, silver impregnation methods were used to localize the preterminal degeneration of fibers in transverse sections of the brainstem 5 and 7 days after CN ablation. CN efferents projected heavily to the lateral superior olive (LSO) ipsilaterally, the medial superior olive (MSO) bilaterally, and contralaterally to the medial (MNTB) and ventral (VNTB) nuclei of the trapezoid body, the ventral (VNLL) and intermediate nuclei of the lateral lemniscus and the central nucleus of the inferior colliculus (ICc). There were smaller projections to the lateral nucleus of the trapezoid body ipsilaterally, the dorsal and dorsomedial periolivary nuclei bilaterally, and the dorsal nucleus of the lateral lemniscus contralaterally. There were sparse projections to the VNLL and ICc ipsilaterally and the CN contralaterally, and a very sparse projection to the contralateral LSO. To determine if CN efferents were glutamatergic and/or aspartatergic, the fresh brainstem was sectioned transversely and samples of the LSO, MSO, MNTB, VNLL, and ICc were taken to measure the electrically evoked release and the uptake of D-[3H]Asp and [14C]Gly or [14C]GABA 3-5 days after the CN ablation. The release studies suggest that only certain of the histologically identified projections ascending from the CN may be glutamatergic and/or aspartatergic. CN ablation depressed D-[3H]Asp release in the MSO bilaterally and in the contralateral MNTB and VNLL, suggesting that the CN efferents to these nuclei may use glutamate or aspartate as a transmitter. It was unclear whether a marginal depression of D-[3H]Asp release in the ipsilateral LSO reflected the presence of glutamatergic CN projections to this nucleus. D-[3H]Asp release in the ICc was unaffected, suggesting that CN efferents to this nucleus may not be glutamatergic. There were no deficits in D-[3H]Asp uptake. [14C]Gly release from the LSO and MSO was unchanged. [14C]Gly uptake was unchanged in the MSO and depressed only in the contralateral LSO, possibly reflecting subnormal uptake activity in endings contributed by contralateral MNTB cells that had lost their CN efferents.(ABSTRACT TRUNCATED AT 400 WORDS)

Azimuthal receptive fields are shaped by GABAergic inhibition in the inferior colliculus of the mustache bat.

1. In this study we examine the effects of GABAergic inhibition on the response properties and the constructed azimuthal receptive fields of 54 excitatory/inhibitory (EI) neurons tuned to 60 kHz in the inferior colliculus of the mustache bat. The constructed azimuthal receptive fields predict the spike counts that would be evoked by different intensities of 60-kHz sounds presented from each of 13 azimuthal locations in the frontal sound field. 2. Action potentials were recorded with a micropipette attached to a multibarrel glass electrode. Bicuculline, an antagonist specific for gamma-aminobutyric acid-A (GABAA) receptors, was iontophoretically applied through the multibarrel electrode. Both monaural and binaural response properties were initially recorded at a variety of interaural intensity disparities (IIDs) and absolute intensities, and the same response properties were subsequently assessed while GABAergic inhibition was blocked by bicuculline. Azimuthal receptive fields both before and during the application of bicuculline were constructed from response properties obtained with earphones after correcting for the directional properties of the ear and the IIDs generated by 60-kHz sounds presented from a variety of azimuthal locations. 3. Bicuculline had virtually no effect on either the monaural or binaural properties of 19 cells (35%). The constructed azimuthal receptive fields of these cells were also unaffected by bicuculline. Presumably the properties of these cells were formed in a lower nucleus, most likely the contralateral lateral superior olive (LSO), and were imposed on the collicular cell via the crossed projection from the LSO to the inferior colliculus, which is known to be excitatory. 4. In more than half of the neurons (65%) GABAergic inhibition influenced one or more features of the cell's response properties and thus its azimuthal receptive field. Some response properties were formed in the colliculus through GABAergic inhibition, whereas others appear to have been shaped initially in a lower nucleus and then further modified by GABAergic inhibition in the inferior colliculus. Moreover, a number of features of GABAergic inhibition that acted on inferior collicular cells were evoked by stimulation of the contralateral (excitatory) ear, whereas other features were influenced by stimulation of the ipsilateral (inhibitory) ear. 5. In 20 cells (37%) blocking GABAergic inhibition reduced or abolished the inhibition evoked by the ipsilateral ear. The receptive fields of cells in which the ipsilaterally evoked inhibition was reduced by bicuculline expanded further into the ipsilateral sound field than they did before bicuculline.(ABSTRACT TRUNCATED AT 400 WORDS)