Lateral Superior Olive Neurons Research Articles

Enhanced Excitatory Connectivity and Disturbed Sound Processing in the Auditory Brainstem of Fragile X Mice.

Hypersensitivity to sounds is one of the prevalent symptoms in individuals with Fragile X syndrome (FXS). It manifests behaviorally early during development and is often used as a landmark for treatment efficacy. However, the physiological mechanisms and circuit-level alterations underlying this aberrant behavior remain poorly understood. Using the mouse model of FXS (Fmr1 KO), we demonstrate that functional maturation of auditory brainstem synapses is impaired in FXS. Fmr1 KO mice showed a greatly enhanced excitatory synaptic input strength in neurons of the lateral superior olive (LSO), a prominent auditory brainstem nucleus, which integrates ipsilateral excitation and contralateral inhibition to compute interaural level differences. Conversely, the glycinergic, inhibitory input properties remained unaffected. The enhanced excitation was the result of an increased number of cochlear nucleus fibers converging onto one LSO neuron, without changing individual synapse properties. Concomitantly, immunolabeling of excitatory ending markers revealed an increase in the immunolabeled area, supporting abnormally elevated excitatory input numbers. Intrinsic firing properties were only slightly enhanced. In line with the disturbed development of LSO circuitry, auditory processing was also affected in adult Fmr1 KO mice as shown with single-unit recordings of LSO neurons. These processing deficits manifested as an increase in firing rate, a broadening of the frequency response area, and a shift in the interaural level difference function of LSO neurons. Our results suggest that this aberrant synaptic development of auditory brainstem circuits might be a major underlying cause of the auditory processing deficits in FXS.SIGNIFICANCE STATEMENT Fragile X Syndrome (FXS) is the most common inheritable form of intellectual impairment, including autism. A core symptom of FXS is extreme sensitivity to loud sounds. This is one reason why individuals with FXS tend to avoid social interactions, contributing to their isolation. Here, a mouse model of FXS was used to investigate the auditory brainstem where basic sound information is first processed. Loss of the Fragile X mental retardation protein leads to excessive excitatory compared with inhibitory inputs in neurons extracting information about sound levels. Functionally, this elevated excitation results in increased firing rates, and abnormal coding of frequency and binaural sound localization cues. Imbalanced early-stage sound level processing could partially explain the auditory processing deficits in FXS.

Microsecond interaural time differences of acoustic transients are decoded by inhibitory-excitatory interactions in neurons of the lateral superior olive

The lateral superior olive (LSO) in the auditory brainstem generates sensitivity to interaural level differences (ILD), an important cue for sound localization, by comparing excitatory (E) input from the ipsilateral ear with inhibitory (I) input from the contralateral ear. Large axosomatic synapses (e.g., calyx of Held) point to the importance of precise temporal processing, but that is not easily reconciled with ILD detection. We propose that the IE interaction allows detection of interaural time differences (ITD) of acoustic transients, to which humans are exquisitely sensitive. We obtained in vivo whole cell recordings of LSO and MSO neurons in the gerbil, while presenting monaural and binaural clicks. We found that ITD functions to clicks in the LSO are surprisingly steep, in contrast to MSO neurons, which are considered the main ITD detectors. Intracellular LSO recordings show EPSPs generated by the ipsilateral click and IPSPs by the contralateral click, where IPSPs often arrive earlier. Binaural spiking is maximally suppressed when the EPSP coincides with the falling slope rather than the peak of the IPSP. We conclude that LSO neurons are more sensitive to ITDs of transients than MSO neurons. This clarifies the importance of timing specializations in the LSO circuit.

Pb exposure prolongs the time period for postnatal transient uptake of 5-HT by murine LSO neurons

Pb exposure is associated with cognitive deficits including Attention Deficit Hyperactivity Disorder (ADHD) in children and alters auditory temporal processing in humans and animals. Serotonin has been implicated in auditory temporal processing and previous studies from our laboratory have demonstrated that developmental Pb decreases expression of serotonin (5-HT) in the adult murine lateral superior olive (LSO). During development, certain non-serotonergic sensory neurons, including auditory LSO neurons, transiently take up 5-HT through the serotonin reuptake transporter (SERT). The uptake of 5-HT is important for development of sensory systems. This study examines the effect of Pb on the serotonergic system in the LSO of the early postnatal mouse. Mice were exposed to moderate Pb (0.01mM) or high Pb (0.1mM) throughout gestation and postnatal day 4 (P4) and P8. We found that Pb exposure prolongs the normal developmental expression of 5-HT by LSO neurons and this is correlated with expression of SERT on LSO cell bodies. The prolonged expression of 5-HT by postnatal LSO neurons is correlated with decreased synaptic immunolabeling within the LSO. This Pb-associated decrease in synaptic density within the LSO could contribute to the auditory temporal processing deficits and cognitive deficits associated with developmental Pb exposure.

Roles for Coincidence Detection in Coding Amplitude-Modulated Sounds.

Many sensory neurons encode temporal information by detecting coincident arrivals of synaptic inputs. In the mammalian auditory brainstem, binaural neurons of the medial superior olive (MSO) are known to act as coincidence detectors, whereas in the lateral superior olive (LSO) roles of coincidence detection have remained unclear. LSO neurons receive excitatory and inhibitory inputs driven by ipsilateral and contralateral acoustic stimuli, respectively, and vary their output spike rates according to interaural level differences. In addition, LSO neurons are also sensitive to binaural phase differences of low-frequency tones and envelopes of amplitude-modulated (AM) sounds. Previous physiological recordings in vivo found considerable variations in monaural AM-tuning across neurons. To investigate the underlying mechanisms of the observed temporal tuning properties of LSO and their sources of variability, we used a simple coincidence counting model and examined how specific parameters of coincidence detection affect monaural and binaural AM coding. Spike rates and phase-locking of evoked excitatory and spontaneous inhibitory inputs had only minor effects on LSO output to monaural AM inputs. In contrast, the coincidence threshold of the model neuron affected both the overall spike rates and the half-peak positions of the AM-tuning curve, whereas the width of the coincidence window merely influenced the output spike rates. The duration of the refractory period affected only the low-frequency portion of the monaural AM-tuning curve. Unlike monaural AM coding, temporal factors, such as the coincidence window and the effective duration of inhibition, played a major role in determining the trough positions of simulated binaural phase-response curves. In addition, empirically-observed level-dependence of binaural phase-coding was reproduced in the framework of our minimalistic coincidence counting model. These modeling results suggest that coincidence detection of excitatory and inhibitory synaptic inputs is essential for LSO neurons to encode both monaural and binaural AM sounds.

Excitation by Axon Terminal GABA Spillover in a Sound Localization Circuit.

Synapses from neurons of the medial nucleus of the trapezoid body (MNTB) onto neurons of the lateral superior olive (LSO) in the auditory brainstem are glycinergic in maturity, but also GABAergic and glutamatergic in development. The role for this neurotransmitter cotransmission is poorly understood. Here we use electrophysiological recordings in brainstem slices from P3-P21 mice to demonstrate that GABA release evoked from MNTB axons can spill over to neighboring MNTB axons and cause excitation by activating GABAAR. This spillover excitation generates patterns of staggered neurotransmitter release from different MNTB axons resulting in characteristic "doublet" postsynaptic currents in LSO neurons. Postembedding immunogold labeling and electron microscopy provide evidence that GABAARs are localized at MNTB axon terminals. Photolytic uncaging of p-hydroxyphenacyl (pHP) GABA demonstrates backpropagation of GABAAR-mediated depolarizations from MNTB axon terminals to the soma, some hundreds of microns away. These somatic depolarizations enhanced somatic excitability by increasing the probability of action potential generation. GABA spillover excitation between MNTB axon terminals may entrain neighboring MNTB neurons, which may play a role in the developmental refinement of the MNTB-LSO pathway. Axonal spillover excitation persisted beyond the second postnatal week, suggesting that this mechanism may play a role in sound localization, by providing new avenues of communication between MNTB neurons via their distal axonal projections. Significance statement: In this study, a new mechanism of neuronal communication between auditory synapses in the mammalian sound localization pathway is described. Evidence is provided that the inhibitory neurotransmitter GABA can spill over between axon terminals to cause excitation of nearby synapses to further stimulate neurotransmitter release. Excitatory GABA spillover between inhibitory axon terminals may have important implications for the development and refinement of this auditory circuit and may play a role in the ability to precisely localize sound sources.

Two-channel models of medial and superior olive based on psychoacoustics

Two nuclei in each hemisphere, the medial superior olive (MSO) and the lateral superior olive (LSO), are responsible for decoding interaural time difference (ITD) and interaural level difference (ILD), respectively [1]. This allows us to localize sound sources in the horizontal plane. Neurons in the MSO are usually modeled as coincidence detectors [1]. Two main approaches to model the MSO appeared during last decades. The most utilized approach (introduced by Jeffress [2]) uses a delay line together with coincidence detectors. The model assumes that the individual neurons in the MSO are sensitive to the different ITD. Another approach (introduced by von Bekesy [3]) accounts for two channels in the brain competing with each other. The concept of two channel model was further developed by Dietz [4] (called rate-code model), and Pulkki and Hirvonen [5] (called count-comparison model), who incorporated new physiological findings. The LSO neurons are consistently reported as an Excitation-Inhibition type [1] and mainly ILD sensitive, thus it can be modeled by a simple subtracter. We present phenomenological models mimicking the functions of the MSO and LSO. The models incorporate the latest physiological findings. However, the models were not designed to simulate responses of the neurons in the MSO and LSO. Instead, the models give data directly comparable with results of subjective lateralization experiments. To simplify the readability, we call these models according to the nuclei which they are mimicking, the MSO and the LSO model. As a front-end, a model of peripheral ear (taken from the literature) is utilized. The MSO model is Excitation-Inhibition type, in principle similar to the two channels, rate-code or the count-comparison models. The LSO model is Excitation-Inhibition type with a simple subtracting unit simulating the process inside the LSO. There is one MSO and LSO model for each hemisphere. Outputs from both hemispheres are compared following the two channels principle. Interaction between the MSO and the LSO model output is not yet possible in this implementation. We compare the outputs of the designed models with the results of psychoacoustic experiments showing the lateralization of pure tones and narrow band noises. The data illustrating the lateralization of pure tones with ITD or ILD was reproduced from the literature [6]. A psychoacoustic experiment was conducted to measure the subjective lateralization of a narrowband noise (central frequency 380 and 760 Hz, bandwidth 1 ERB) with interaural phase or level differences. Seven and five normal hearing listeners participated in the case of interaural phase and level differences test, respectively. A good agreement between the model data and subjective lateralization was observed.

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.

Linear coding of complex sound spectra by discharge rate in neurons of the medial nucleus of the trapezoid body (MNTB) and its inputs

The interaural level difference (ILD) cue to sound location is first encoded in the lateral superior olive (LSO). ILD sensitivity results because the LSO receives excitatory input from the ipsilateral cochlear nucleus and inhibitory input indirectly from the contralateral cochlear nucleus via glycinergic neurons of the ipsilateral medial nucleus of the trapezoid body (MNTB). It is hypothesized that in order for LSO neurons to encode ILDs, the sound spectra at both ears must be accurately encoded via spike rate by their afferents. This spectral-coding hypothesis has not been directly tested in MNTB, likely because MNTB neurons have been mostly described and studied recently in regards to their abilities to encode temporal aspects of sounds, not spectral. Here, we test the hypothesis that MNTB neurons and their inputs from the cochlear nucleus and auditory nerve code sound spectra via discharge rate. The Random Spectral Shape (RSS) method was used to estimate how the levels of 100-ms duration spectrally stationary stimuli were weighted, both linearly and non-linearly, across a wide band of frequencies. In general, MNTB neurons, and their globular bushy cell inputs, were found to be well-modeled by a linear weighting of spectra demonstrating that the pathways through the MNTB can accurately encode sound spectra including those resulting from the acoustical cues to sound location provided by head-related directional transfer functions (DTFs). Together with the anatomical and biophysical specializations for timing in the MNTB-LSO complex, these mechanisms may allow ILDs to be computed for complex stimuli with rapid spectrotemporally-modulated envelopes such as speech and animal vocalizations and moving sound sources.

Development of glycinergic innervation to the murine LSO and SPN in the presence and absence of the MNTB.

Neurons in the superior olivary complex (SOC) integrate excitatory and inhibitory inputs to localize sounds in space. The majority of these inhibitory inputs have been thought to arise within the SOC from the medial nucleus of the trapezoid body (MNTB). However, recent work demonstrates that glycinergic innervation of the SOC persists in Egr2; En1CKO mice that lack MNTB neurons, suggesting that there are other sources of this innervation (Jalabi et al., 2013). To study the development of MNTB- and non-MNTB-derived glycinergic SOC innervation, we compared immunostaining patterns of glycine transporter 2 (GlyT2) at several postnatal ages in control and Egr2; En1CKO mice. GlyT2 immunostaining was present at birth (P0) in controls and reached adult levels by P7 in the superior paraolivary nucleus (SPN) and by P12 in the lateral superior olive (LSO). In Egr2; En1CKO mice, glycinergic innervation of the LSO developed at a similar rate but was delayed by one week in the SPN. Conversely, consistent reductions in the number of GlyT2+ boutons located on LSO somata were seen at all ages in Egr2; En1CKO mice, while these numbers reached control levels in the SPN by adulthood. Dendritic localization of GlyT2+ boutons was unaltered in both the LSO and SPN of adult Egr2; En1CKO mice. On the postsynaptic side, adult Egr2; En1CKO mice had reduced glycine receptor α1 (GlyRα1) expression in the LSO but normal levels in the SPN. GlyRα2 was not expressed by LSO or SPN neurons in either genotype. These findings contribute important information for understanding the development of MNTB- and non-MNTB-derived glycinergic pathways to the mouse SOC.

On the precision of neural computation with interaural level differences in the lateral superior olive

Interaural level difference (ILD) is one of the basic binaural clues in the spatial localization of a sound source. Due to the acoustic shadow cast by the head, a sound source out of the medial plane results in an increased sound level at the nearer ear and a decreased level at the distant ear. In the mammalian auditory brainstem, the ILD is processed by a neuronal circuit of binaural neurons in the lateral superior olive (LSO). These neurons receive major excitatory projections from the ipsilateral side and major inhibitory projections from the contralateral side. As the sound level is encoded predominantly by the neuronal discharge rate, the principal function of LSO neurons is to estimate and encode the difference between the discharge rates of the excitatory and inhibitory inputs. Two general mechanisms of this operation are biologically plausible: (1) subtraction of firing rates integrated over longer time intervals, and (2) detection of coincidence of individual spikes within shorter time intervals. However, the exact mechanism of ILD evaluation is not known. Furthermore, given the stochastic nature of neuronal activity, it is not clear how the circuit achieves the remarkable precision of ILD assessment observed experimentally. We employ a probabilistic model and complementary computer simulations to investigate whether the two general mechanisms are capable of the desired performance. Introducing the concept of an ideal observer, we determine the theoretical ILD accuracy expressed by means of the just-noticeable difference (JND) in dependence on the statistics of the interacting spike trains, the overall firing rate, detection time, the number of converging fibers, and on the neural mechanism itself. We demonstrate that the JNDs rely on the precision of spike timing; however, with an appropriate parameter setting, the lowest theoretical values are similar or better than the experimental values. Furthermore, a mechanism based on excitatory and inhibitory coincidence detection may give better results than the subtraction of firing rates.This article is part of a Special Issue entitled Neural Coding 2012.

Long term depression of MNTB-LSO synapses is expressed postsynaptically in developing circling mice

Early onset long term depression (LTD) during the first postnatal week has rarely been demonstrated at the medial nucleus of trapezoid body (MNTB) – lateral superior olive (LSO) synapses in spite of many favorable conditions, such as depolarizing synapses and glutamate co-release from MNTB terminals. Thus, we tested the early expression of LTD at MNTB-LSO synapses during the first postnatal week using circling mice, whose main transmitter is glutamate at MNTB-LSO synapses. Tetanic stimulation on MNTB elicited LTD of postsynaptic currents recorded at LSO neurons in P0–P3 homozygous (cir/cir) mice (45.8±0.3% of the control, n=7) and heterozygous (+/cir) mice (43.3±0.4% of the control, n=7). The magnitude of LTD decreased in P8–P12 heterozygous (+/cir) mice (84.5±0.3% of the control, n=7), but was maintained in P8–P12 homozygous (cir/cir) mice (38.2±0.3% of the control, n=9). Glutamatergic LTD observed in homozygous (cir/cir) mice and glycinergic LTD observed heterozygous (+/cir) mice showed similar pattern of change. As currents induced by the pressure application of glycine on LSO neurons were reduced by tetanic stimulation in P0–P3 heterozygous (+/cir) mice, LTD was thought to occur at postsynaptic sites. Our results suggest that LTD might occur in vivo and participate in the synaptic silencing and strengthening of MNTB-LSO synapses, which is most active during the first postnatal week.

Embryonic assembly of auditory circuits: spiral ganglion and brainstem

During early development, peripheral sensory systems generate physiological activity prior to exposure to normal environmental stimuli. This activity is thought to facilitate maturation of these neurons and their connections, perhaps even promoting efficacy or modifying downstream circuitry. In the mammalian auditory system, initial connections form at embryonic ages, but the functional characteristics of these early neural connections have not been assayed. We investigated processes of embryonic auditory development using a whole-head slice preparation that preserved connectivity between peripheral and brainstem stations of the auditory pathway. Transgenic mice expressing fluorescent protein provided observation of spiral ganglion and cochlear nucleus neurons to facilitate targeted electrophysiological recording. Here we demonstrate an apparent peripheral-to-central order for circuit maturation. Spiral ganglion cells acquire action potential-generating capacity at embryonic day 14 (E14), the earliest age tested, and action potential waveforms begin to mature in advance of comparable states for neurons of the ventral cochlear nucleus (VCN) and medial nucleus of the trapezoid body (MNTB). In accordance, auditory nerve synapses in the VCN are functional at E15, prior to VCN connectivity with the MNTB, which occurs at least 1 day later. Spiral ganglion neurons exhibit spontaneous activity at least by E14 and are able to drive third-order auditory brainstem neurons by E17. This activity precedes cochlear-generated wave activity by 4 days and ear canal opening by at least 2 weeks. Together, these findings reveal a previously unknown initial developmental phase for auditory maturation, and further implicate the spiral ganglion as a potential controlling centre in this process.

Repertoire of high voltage-activated Ca2+ channels in the lateral superior olive: functional analysis in wild-type, Cav1.3−/−, and Cav1.2DHP−/− mice

Voltage-gated Ca(2+) (Ca(v))1.3 α-subunits of high voltage-activated Ca(2+) channels (HVACCs) are essential for Ca(2+) influx and transmitter release in cochlear inner hair cells and therefore for signal transmission into the central auditory pathway. Their absence leads to deafness and to striking structural changes in the auditory brain stem, particularly in the lateral superior olive (LSO). Here, we analyzed the contribution of various types of HVACCs to the total Ca(2+) current (I(Ca)) in developing mouse LSO neurons to address several questions: do LSO neurons express functional Ca(v)1.3 channels? What other types of HVACCs are expressed? Are there developmental changes? Do LSO neurons of Ca(v)1.3(-/-) mice show any compensatory responses, namely, upregulation of other HVACCs? Our electrophysiological and pharmacological results showed the presence of functional Ca(v)1.3 and Ca(v)1.2 channels at both postnatal days 4 and 12. Aside from these L-type channels, LSO neurons also expressed functional P/Q-type, N-type, and, most likely, R-type channels. The relative contribution of the four different subtypes to I(Ca) appeared to be 45%, 29%, 22%, and 4% at postnatal day 12, respectively. The physiological results were flanked and extended by quantitative RT-PCR data. Altogether, LSO neurons displayed a broad repertoire of HVACC subtypes. Genetic ablation of Ca(v)1.3 resulted in functional reorganization of some other HVACCs but did not restore normal I(Ca) properties. Together, our results suggest that several types of HVACCs are of functional relevance for the developing LSO. Whether on-site loss of Ca(v)1.3, i.e., in LSO neurons, contributes to the recently described malformation of the LSO needs to be determined by using tissue-specific Ca(v)1.3(-/-) animals.

Low-threshold potassium currents stabilize IID-sensitivity in the inferior colliculus

The inferior colliculus (IC) is a midbrain nucleus that exhibits sensitivity to differences in interaural time and intensity (ITDs and IIDs) and integrates information from the auditory brainstem to provide an unambiguous representation of sound location across the azimuth. Further upstream, in the lateral superior olive (LSO), absence of low-threshold potassium currents in Kcna1−/− mice interfered with response onset timing and restricted IID-sensitivity to the hemifield of the excitatory ear. Assuming the IID-sensitivity in the IC to be at least partly inherited from LSO neurons, the IC IID-encoding was compared between wild-type (Kcna1+/+) and Kcna1−/− mice. We asked whether the effect observed in the Kcna1−/− LSO is (1) simply propagated into the IC, (2) is enhanced and amplified or, (3) alternatively, is compensated and so no longer detectable. Our results show that general IC response properties as well as the distribution of IID-functions were comparable in Kcna1−/− and Kcna1+/+ mice. In agreement with the literature IC neurons exhibited a higher level-invariance of IID-sensitivity compared to LSO neurons. However, manipulating the timing between the inputs of the two ears caused significantly larger shifts of IID-sensitivity in Kcna1−/− mice, whereas in the wild-type IC the IID functions were stable and less sensitive to changes of the temporal relationship between the binaural inputs. We conclude that the IC not only inherits IID-sensitivity from the LSO, but that the convergence with other, non-olivary inputs in the wild-type IC acts to quality-control, consolidate, and stabilize IID representation; this necessary integration of inputs is impaired in the absence of the low-threshold potassium currents mediated by Kv1.1.

Decreased Immunoreactivities of the Chloride Transporters, KCC2 and NKCC1, in the Lateral Superior Olive Neurons of Kanamycin-treated Rats

ObjectivesFrom our previous study about the weak expressions of potassium-chloride (KCC2) and sodium-potassium-2 chloride (NKCC1) co-transporters in the lateral superior olive (LSO) in circling mice, we hypothesized that partially damaged cochlea of circling mice might be a cause of the weak expressions of KCC2 or NKCC1. To test this possibility, we reproduced the altered expressions of KCC2 and NKCC1 in the LSO of rats, whose cochleae were partially destroyed with kanamycin.MethodsRat pups were treated with kanamycin from postnatal (P)3 to P8 (700 mg/kg, subcutaneous injection, twice a day) and sacrificed for immunohistochemical analysis, scanning electron microscope (SEM) and auditory brain stem response.ResultsThe SEM study revealed partially missing hair cells in P9 rats treated with kanamycin, and the hearing threshold was elevated to 63.8±2.5 dB SPL (4 ears) at P16. Both KCC2 and NKCC1 immunoreactivities were more prominent in control rats on P16. On 9 paired slices, the mean densities of NKCC1 immunoreactivities were 118.0±1.0 (control) and 112.2±1.2 (kanamycin treated), whereas those of KCC2 were 115.7±1.5 (control) and 112.0±0.8 (kanamycin treated).ConclusionWe concluded that weak expressions of KCC2 and NKCC1 in circling mice were due to partial destruction of cochleae.

A Modeling Study of the Responses of the Lateral Superior Olive to Ipsilateral Sinusoidally Amplitude-Modulated Tones

The lateral superior olive (LSO) is a brainstem nucleus that is classically understood to encode binaural information in high-frequency sounds. Previous studies have shown that LSO cells are sensitive to envelope interaural time difference in sinusoidally amplitude-modulated (SAM) tones (Joris and Yin, J Neurophysiol 73:1043–1062, 1995; Joris, J Neurophysiol 76:2137–2156, 1996) and that a subpopulation of LSO neurons exhibit low-threshold potassium currents mediated by Kv1 channels (Barnes-Davies et al., Eur J Neurosci 19:325–333, 2004). It has also been shown that in many LSO cells the average response rate to ipsilateral SAM tones decreases with modulation frequency above a few hundred Hertz (Joris and Yin, J Neurophysiol 79:253–269, 1998). This low-pass feature is not directly inherited from the inputs to the LSO since the response rate of these input neurons changes little with increasing modulation frequency. In the current study, an LSO cell model is developed to investigate mechanisms consistent with the responses described above, notably the emergent rate decrease with increasing frequency. The mechanisms explored included the effects of after-hyperpolarization (AHP) channels, the dynamics of low-threshold potassium channels (KLT), and the effects of background inhibition. In the model, AHP channels alone were not sufficient to induce the observed rate decrease at high modulation frequencies. The model also suggests that the background inhibition alone, possibly from the medial nucleus of the trapezoid body, can account for the small rate decrease seen in some LSO neurons, but could not explain the large rate decrease seen in other LSO neurons at high modulation frequencies. In contrast, both the small and large rate decreases were replicated when KLT channels were included in the LSO neuron model. These results support the conclusion that KLT channels may play a major role in the large rate decreases seen in some units and that background inhibition may be a contributing factor, a factor that could be adequate for small decreases.

Comparative posthearing development of inhibitory inputs to the lateral superior olive in gerbils and mice

Interaural intensity differences are analyzed in neurons of the lateral superior olive (LSO) by integration of an inhibitory input from the medial nucleus of the trapezoid body (MNTB), activated by sound from the contralateral ear, with an excitatory input from the ipsilateral cochlear nucleus. The early postnatal refinement of this inhibitory MNTB-LSO projection along the tonotopic axis of the LSO has been extensively studied. However, little is known to what extent physiological changes at these inputs also occur after the onset of sound-evoked activity. Using whole-cell patch-clamp recordings of LSO neurons in acute brain stem slices, we analyzed the developmental changes of inhibitory synaptic currents evoked by MNTB fiber stimulation occurring after hearing onset. We compared these results in gerbils and mice, two species frequently used in auditory research. Our data show that neither the number of presumed input fibers nor the conductance of single fibers significantly changed after hearing onset. Also the amplitude of miniature inhibitory currents remained constant during this developmental period. In contrast, the kinetics of inhibitory synaptic currents greatly accelerated after hearing onset. We conclude that tonotopic refinement of inhibitory projections to the LSO is largely completed before the onset of hearing, whereas acceleration of synaptic kinetics occurs to a large part after hearing onset and might thus be dependent on proper auditory experience. Surprisingly, inhibitory input characteristics, as well as basic membrane properties of LSO neurons, were rather similar in gerbils and mice.

Paired recordings from distant inhibitory neuron pairs by a sequential scanning approach

Simultaneous recordings from connected neuron pairs have brought important insights into synaptic communication between neurons. However, patch clamp recordings from neuron pairs have been largely restricted to brain areas in which connections among nearby neurons exist at a relatively high probability. In the case of more distant connections or in areas in which neurons are connected with low probability, recordings from synaptically connected neuron pairs have remained scarce. Here, we present a method that allows dual recordings from remotely connected neuron pairs by scanning potential presynaptic neurons. The applicability of this new approach was tested in the inhibitory pathway from the medial nucleus of the trapezoid body (MNTB) to the lateral superior olive (LSO), a sound localization pathway in the auditory brainstem. Using a three-step approach that sequentially combines focal uncaging of glutamate, pressure application of glutamate, and loose patch recordings allowed us to reliably achieve recordings from distant, synaptically connected GABA/glycinergic MNTB–LSO neuron pairs. Our results demonstrate that single MNTB neurons evoke highly variable mono-synaptic responses in developing LSO neurons, and heterogeneous short term synaptic dynamics, suggesting local variations in the refinement of these inhibitory connections. Paired recordings, enabled by scanning of remotely connected pairs, will be highly useful to perform detailed investigations of the synaptic function and plasticity from these circuits during the period of developmental refinement. In general, this method should provide a valuable tool to find connected neurons in other brain areas in which recording from candidate pairs has a low success rate.

Cav1.3 Calcium Channels Are Required for Normal Development of the Auditory Brainstem

Within the Ca(v)1 family of voltage-gated calcium channels, Ca(v)1.2 and Ca(v)1.3 channels are the predominant subtypes in the brain. Whereas specific functions for each subtype were described in the adult brain, their role in brain development is poorly understood. Here we assess the role of Ca(v)1.3 subunits in the activity-dependent development of the auditory brainstem. We used Ca(v)1.3-deficient (Ca(v)1.3(-/-)) mice because these mice lack cochlea-driven activity that deprives the auditory centers from peripheral input. We found a drastically reduced volume in all auditory brainstem centers (range 25-59%, total 35%), which was manifest before hearing onset. A reduction was not obvious outside the auditory system. The lateral superior olive (LSO) was strikingly malformed in Ca(v)1.3(-/-) mice and had fewer neurons (1/3 less). The remaining LSO neurons displayed normal dendritic trees and received functional glutamatergic input, yet they fired action potentials predominantly with a multiple pattern upon depolarization, in contrast to the single firing pattern prevalent in controls. The latter finding appears to be due to a reduction of dendrototoxin-sensitive potassium conductances, presumably mediated through the K(v)1.2 subtype. Fura2 imaging provided evidence for functional Ca(v)1.3 channels in the LSO of wild-type mice. Our results imply that Ca(v)1.3 channels are indispensable for the development of the central auditory system. We propose that the unique LSO phenotype in Ca(v)1.3(-/-) mice, which hitherto was not described in other hereditary deafness models, is caused by the synergistic contribution of two factors: on-site loss of Ca(v)1.3 channels in the neurons plus lack of peripheral input.

Neural sensitivity to interaural level differences determines virtual acoustic space minimum audible angles for single neurons in the lateral superior olive.

The minimum audible angle (MAA), the smallest angle separating two sound sources that can be reliably discriminated, is a psychophysical measure of spatial acuity. In humans and cats, MAAs for tone and noise stimuli range from 1–5 deg. For high-frequency (>1.5 kHz) tones the predominant cue for azimuth is the interaural level difference (ILD). Neurophysiologically, ILDs are first encoded in the lateral superior olive (LSO). Here, we examined the ability of LSO neurons in cats to signal changes in the azimuth of noise sources. Using measurements of head related transfer functions, the virtual acoustic space technique was used to manipulate source azimuth in the physiological experiments. For each neuron detection theory was used to compute the smallest increment in azimuth necessary to discriminate that change based on discharge rate and response variability. Minimum neural MAAs were 2.3 deg for midline sources (median = 4.5 deg, n = 32 neurons). The good neural acuity for spatial location will be explained in terms of the changes in the acoustic ILD cue with changes in source location along with the underlying sensitivity of the LSO neurons to the ILD cue. LSO neurons can signal changes in sound azimuth that match or exceeded behavioral capabilities. [NIH DC006865.]