Mammalian Auditory System Research Articles

Does age protect against loss of tonotopy after acute deafness in adulthood?

The mammalian auditory system develops a topographical representation of sound frequencies along its pathways, also called tonotopy. In contrast, sensory deprivation during early development results in no or only rudimentary tonotopic organization. This study addresses two questions: (1) How robust is the central tonotopy when hearing fails in adulthood? (2) What role does age play at time of deafness? To address these questions, we deafened young and old adult rats with previously normal hearing. One month after deafening, both groups were unilaterally supplied with cochlear implants and electrically stimulated for 2 h. The central auditory neurons, which were activated as a result of the local electrical intracochlear stimulation, were visualized using Fos staining. While the auditory system of young rats lost the tonotopic organization throughout the brainstem, the auditory system of the older rats mainly sustained its tonotopy. It can be proposed that plasticity prevails in the central auditory system of young adult rats, while network stability prevails in the brains of aging rats. Consequently, age may be an important factor in protecting a hearing-experienced adult auditory system from a rapid loss of tonotopy when suffering from acute hearing loss. Furthermore, the study provides compelling evidence that acute deafness in young adult patients should be diagnosed as early as possible to prevent maladaptation of the central auditory system and thus achieve the optimal hearing outcome with a hearing prosthesis.

3D reconstruction of the mouse cochlea from scRNA-seq data suggests morphogen-based principles in apex-to-base specification

In the mammalian auditory system, frequency discrimination depends on numerous morphological and physiological properties of the organ of Corti, which gradually change along the apex-to-base (tonotopic) axis of the organ. For example, the basilar membrane stiffness changes tonotopically, thus affecting the tuning properties of individual hair cells. At the molecular level, those frequency-specific characteristics are mirrored by gene expression gradients; however, the molecular mechanisms controlling tonotopic gene expression in the mouse cochlea remain elusive. Through analyzing single-cell RNA sequencing (scRNA-seq) data from E12.5 and E14.5 time points, we predicted that morphogens, rather than a cell division-associated mechanism, confer spatial identity in the extending cochlea. Subsequently, we reconstructed the developing cochlea in 3D space from scRNA-seq data to investigate the molecular pathways mediating positional information. The retinoic acid (RA) and hedgehog pathways were found to form opposing apex-to-base gradients, and functional interrogation using mouse cochlear explants suggested that both pathways jointly specify the longitudinal axis.

The role of GABAB receptors in the subcortical pathways of the mammalian auditory system.

GABAB receptors are G-protein coupled receptors for the inhibitory neurotransmitter GABA. Functional GABAB receptors are formed as heteromers of GABAB1 and GABAB2 subunits, which further associate with various regulatory and signaling proteins to provide receptor complexes with distinct pharmacological and physiological properties. GABAB receptors are widely distributed in nervous tissue, where they are involved in a number of processes and in turn are subject to a number of regulatory mechanisms. In this review, we summarize current knowledge of the cellular distribution and function of the receptors in the inner ear and auditory pathway of the mammalian brainstem and midbrain. The findings suggest that in these regions, GABAB receptors are involved in processes essential for proper auditory function, such as cochlear amplifier modulation, regulation of spontaneous activity, binaural and temporal information processing, and predictive coding. Since impaired GABAergic inhibition has been found to be associated with various forms of hearing loss, GABAB dysfunction could also play a role in some pathologies of the auditory system.

ALFONSO CORTI: L’UOMO, LA VITA, LE OPERE

ABSTRACT. – Marquis Alfonso Corti di Santo Stefano Belbo, descendant from a rich and noble family of Lombardy and Piedmont, described for the first time, in 1851, a series of cellular structures of the inner ear, basic for the auditory function, now known as “Corti’s organ”. Corti, born in Gambarana near Pavia on June 25, 1822, enrolled in 1841as a medical student at the University of Pavia. Here his favorite study was microanatomy, under professors Bartolomeo Panizza and Mauro Rusconi. In 1845, Corti moved to Vienna to complete his medical studies and to work at the Anatomical institute of the University under the supervision of professor Joseph Hyrtl and in August 1847 received the degree in medicine, with a thesis on the cardiovascular system of a lizard, the Psammosaurus griseus. At the end of December 1847 Corti was chosen as Hyrtl’s second prosector, but he soon had to relinquish this position. With the outbreak of the 1848 Revolution he left Vienna and, after a brief military service in Italy, from February to August 1849 was in Bern, where he began his own-microscopic studies in collaboration with the physiologist Gustav Gabriel Valentin. Except for a trip to England, where he met some important microscopists, Corti spent the rest of 1849 with his relatives in Paris, working at the Institute of Anatomy of the Sorbonne. In the middle of January 1850, Corti left for Wurzburg Institute of Anatomy, directed by Albert Kölliker, where he began to work on the mammalian auditory system. In August 1850 Corti spent a short time at the Observatorium microscopicum in Utrech, then he returned to Würzburg and later to Paris, to complete his study of at least 200 cochleas of man and different animals. His famous paper, “Recherches sur l’organe de l’ouie des mammiferes”, appeared in 1851 in Kölliker’s journal Zeitschrift für wissenschaftliche Zoologie. After the publication Corti left Paris for Turin where he had to interrupt his scientific work in order to deal with the division of his father estate. Suddenly and inexplicably, Corti gave up any scientific activity and disappeared from the scientific world. In 1855 Corti married Maria Bettinzoli, lady of noble birth, who presented him with a daughter Bianca, and a son Gaspare, but in 1861 she died, leaving him with the responsibility of rearing the children. Corti spent the rest of his life as a country nobleman and selected winegrower in his Villa Mazzolino in Corvino San Quirico near Casteggio, soon restricted in his mobility due to a severe arthritis deformans, and died in Corvino on October 2, 1876. After Corti brought his relations with his anatomist colleagues in 1854, his stature began to fade. During the second half of the 19th century, Corti’s Recherches were cited more and more in articles and treatises on otology, but the memory of their author has been lost. Corti passed like a meteor in the scientific world of the mid 19th century and it was only at the beginning of the 20th century that some researches began to wonder about the man behind the name of Corti’s organ.

Bandpass Shape of Distortion-Product Otoacoustic Emission Ratio Functions Reflects Cochlear Frequency Tuning in Normal-Hearing Mice

The frequency selectivity of the mammalian auditory system is critical for discriminating complex sounds like speech. This selectivity derives from the sharp tuning of the cochlea’s mechanical response to sound, which is largely attributed to the amplification of cochlear vibrations by outer hair cells (OHCs). Due to its nonlinearity, the amplification process also leads to the generation of distortion products (DPs), some of which propagate out to the ear canal as DP otoacoustic emissions (DPOAEs). However, the insight that these signals provide about the tuned micro- and macro-mechanics underlying their generation remains unclear. Using optical coherence tomography to measure cochlear vibrations in mice, we show that the cochlea’s frequency tuning is reflected in the bandpass shape that is observed in DPOAE amplitudes when the ratio of the two evoking stimulus frequencies is varied (here termed DPOAE “ratio functions”). The tuning sharpness of DPOAE ratio functions and cochlear vibrations co-varied with stimulus level, with a similar quantitative agreement in tuning sharpness observed for both apical and mid-cochlear locations. Measurement of intracochlear DPs revealed that the tuning of the DPOAE ratio functions was not caused by mechanisms that shape DPs locally near where they are generated. Instead, simple model simulations indicate that the bandpass shape is due to a more global wave interference phenomenon. It appears that the filtering of DPOAEs by wave interactions over an extended spatial region allows them to provide a window onto the frequency tuning of single cochlear locations.

Development of the Hair Cells of the Human Cochlea: A Scanning Electron Microscopic Study.

In the mammalian auditory system, the cochlea is the first to attain structural and functional maturity. Although ultrastructural details of the developing cochlea of lower animals have been elucidated in the last few decades, comprehensive studies on human cochlea are lacking. In the present investigation we studied the development and maturation of the hair cells of ten human fetal cochlea from gestational weeks (GW) 12 to 37 by scanning electron microscopy. We observed undifferentiated hair cells possessing numerous surface projections and long kinocilium during GW 14. At GW16, the primitive hair cells were arranged in one inner and four outer rows and had globular apices indicating the initiation of stereocilia formation. By GW 22, the globular apices were replaced by linear stereocilia and occasional kinocillia. Mature hair cells with sterocilia were observed in the basal turn at 30th week of gestation. At GW 37, the stereocilia were arranged in a typical "V" shaped pattern at the middle and apical coil, while the stereocilia of the basal turn were shorter in length resembling the adult cochlea. The inner hair cells were long and slender while outer hair cells were pear shaped, kinocilium were absent and the tunnel of Corti were well formed. It is concluded that in human, the morphological maturation of the hair cells starts in the basal turn around GW 22 and continues till 37th week in the apical turn indicating that early maturation of the cochlea may have a role on development of the higher auditory pathway connections.

Dog and human neural sensitivity to voicelikeness: A comparative fMRI study

Voice-sensitivity in the auditory cortex of a range of mammals has been proposed to be determined primarily by tuning to conspecific auditory stimuli, but recent human findings indicate a role for a more general tuning to voicelikeness. Vocal emotional valence, a central characteristic of vocalisations, has been linked to the same basic acoustic parameters across species. Comparative neuroimaging revealed that during voice perception, such acoustic parameters modulate emotional valence-sensitivity in auditory cortical regions in both family dogs and humans. To explore the role of voicelikeness in auditory emotional valence-sensitivity across species, here we constructed artificial emotional sounds in two sound categories: voice-like vs. sine-wave sounds, parametrically modulating two main acoustic parameters, f0 and call length. We hypothesised that if mammalian auditory systems are characterised by a general tuning to voicelikeness, voice-like sounds will be processed preferentially, and acoustic parameters for voice-like sounds will be processed differently than for sine-wave sounds - both in dogs and humans. We found cortical areas in both species that responded stronger to voice-like than to sine-wave stimuli, while there were no regions responding stronger to sine-wave sounds in either species. Additionally, we found that in bilateral primary and emotional valence-sensitive auditory regions of both species, the processing of voice-like and sine-wave sounds are modulated by f0 in opposite ways. These results reveal functional similarities between evolutionarily distant mammals for processing voicelikeness and its effect on processing basic acoustic cues of vocal emotions.

The Impact of Multivesicular Release on the Transmission of Sensory Information by Ribbon Synapses.

The statistics of vesicle release determine how synapses transfer information, but the classical Poisson model of independent release does not always hold at the first stages of vision and hearing. There, ribbon synapses also encode sensory signals as events comprising two or more vesicles released simultaneously. The implications of such coordinated multivesicular release (MVR) for spike generation are not known. Here we investigate how MVR alters the transmission of sensory information compared with Poisson synapses using a pure rate-code. We used leaky integrate-and-fire models incorporating the statistics of release measured experimentally from glutamatergic synapses of retinal bipolar cells in zebrafish (both sexes) and compared these with models assuming Poisson inputs constrained to operate at the same average rates. We find that MVR can increase the number of spikes generated per vesicle while reducing interspike intervals and latency to first spike. The combined effect was to increase the efficiency of information transfer (bits per vesicle) over a range of conditions mimicking target neurons of different size. MVR was most advantageous in neurons with short time constants and reliable synaptic inputs, when less convergence was required to trigger spikes. In the special case of a single input driving a neuron, as occurs in the auditory system of mammals, MVR increased information transfer whenever spike generation required more than one vesicle. This study demonstrates how presynaptic integration of vesicles by MVR can increase the efficiency with which sensory information is transmitted compared with a rate-code described by Poisson statistics.SIGNIFICANCE STATEMENT Neurons communicate by the stochastic release of vesicles at the synapse and the statistics of this process will determine how information is represented by spikes. The classical model is that vesicles are released independently by a Poisson process, but this does not hold at ribbon-type synapses specialized to transmit the first electrical signals in vision and hearing, where two or more vesicles can fuse in a single event by a process termed coordinated multivesicular release. This study shows that multivesicular release can increase the number of spikes generated per vesicle and the efficiency of information transfer (bits per vesicle) over a range of conditions found in the retina and peripheral auditory system.

Developmental spontaneous activity promotes formation of sensory domains, frequency tuning and proper gain in central auditory circuits.

Neurons that process sensory information exhibit bursts of electrical activity during development, providing early training to circuits that will later encode similar features of the external world. In the mammalian auditory system, this intrinsically generated activity emerges from the cochlea prior to hearing onset, but its role in maturation of auditory circuitry remains poorly understood. We show that selective suppression of cochlear supporting cell spontaneous activity disrupts patterned burst firing of central auditory neurons without affecting cell survival or acoustic thresholds. However, neurons in the inferior colliculus of these mice exhibit enhanced acoustic sensitivity and broader frequency tuning, resulting in wider isofrequency laminae. Despite this enhanced neural responsiveness, total tone-responsive regions in the auditory cortex are substantially smaller. Thus, disruption of pre-hearing cochlear activity causes profound changes in neural encoding of sound, with important implications for restoration of hearing in individuals who experience reduced activity during this critical developmental period.

A numerical approach to investigating the mechanisms behind tonotopy in the bush-cricket inner-ear.

Bush-crickets (or katydids) have sophisticated and ultrasonic ears located in the tibia of their forelegs, with a working mechanism analogous to the mammalian auditory system. Their inner-ears are endowed with an easily accessible hearing organ, the crista acustica (CA), possessing a spatial organisation that allows for different frequencies to be processed at specific graded locations within the structure. Similar to the basilar membrane in the mammalian ear, the CA contains mechanosensory receptors which are activated through the frequency dependent displacement of the CA. While this tonotopical arrangement is generally attributed to the gradual stiffness and mass changes along the hearing organ, the mechanisms behind it have not been analysed in detail. In this study, we take a numerical approach to investigate this mechanism in the Copiphora gorgonensis ear. In addition, we propose and test the effect of the different vibration transmission mechanisms on the displacement of the CA. The investigation was carried out by conducting finite-element analysis on a three-dimensional, idealised geometry of the C. gorgonensis inner-ear, which was based on precise measurements. The numerical results suggested that (i) even the mildest assumptions about stiffness and mass gradients allow for tonotopy to emerge, and (ii) the loading area and location for the transmission of the acoustic vibrations play a major role in the formation of tonotopy.

Spatial acuity of the bottlenose dolphin (Tursiops truncatus) biosonar system with a bat and human comparison.

Horizontal angular resolution was measured in two bottlenose dolphins using a two-alternative forced-choice, biosonar target discrimination paradigm. The task required a stationary dolphin positioned in a hoop to discriminate two physical targets at a range of 4 m. The angle separating the targets was manipulated to estimate an angular discrimination threshold of 1.5°. In a second experiment, a similar two-target biosonar discrimination task was conducted with one free-swimming dolphin, to test whether its emission beam was a critical factor in discriminating the targets. The spatial separation between two targets was manipulated to measure a discrimination threshold of 6.7 cm. There was a relationship between differences in acoustic signals received at each target and the dolphin's performance. The results of the angular resolution experiment were in good agreement with measures of the minimum audible angle of both dolphins and humans and remarkably similar to measures of angular difference discrimination in echolocating dolphins, bats, and humans. The results suggest that horizontal auditory spatial acuity may be a common feature of the mammalian auditory system rather than a specialized feature exclusive to echolocating auditory predators.

Unloading outer hair cell bundles in vivo does not yield evidence of spontaneous oscillations in the mouse cochlea

Along with outer hair cell (OHC) somatic electromotility as the actuator of cochlear amplification, active hair bundle motility may be a complementary mechanism in the mammalian auditory system. Here, we searched the mouse cochlea for the presence of spontaneous bundle oscillations that have been observed in non-mammalian ears. In those systems, removal of the overlying membrane is necessary for spontaneous bundle oscillations to manifest. Thus, we used a genetic mouse model with a C1509G (cysteine-to-glycine) point mutation in the Tecta gene where the tectorial (TM) is lifted away from the OHC bundles, allowing us to explore whether unloaded bundles spontaneously oscillate. We used VOCTV in vivo to detect OHC length changes due to electromotility as a proxy for the spontaneous opening and closing of the mechanoelectrical transduction (MET) channels associated with bundle oscillation. In wild type mice with the TM attached to OHC bundles, we did find peaks in vibratory magnitude spectra. Such peaks were not observed in the mutants where the TM is detached from the OHC bundles. Statistical analysis of the time signals indicates that these peaks do not signify active oscillations. Rather, they are filtered responses of the sensitive wild type cochlea to weak background noise. We therefore conclude that, to the limits of our system (∼30 pm), there is no spontaneous mechanical activity that manifests as oscillations in OHC electromotility within the mouse cochlea, arguing that unloaded OHC bundles do not oscillate in vivo.This article is part of the Special Issue Outer hair cell Edited by Joseph Santos-Sacchi and Kumar Navaratnam.

A Novel in vitro Model Delineating Hair Cell Regeneration and Neural Reinnervation in Adult Mouse Cochlea.

The study of an adult mammalian auditory system, such as regeneration, has been hampered by the lack of an in vitro system in which hypotheses can be tested efficiently. This is primarily due to the fact that the adult inner ear is encased in the toughest bone of the body, whereas its removal leads to the death of the sensory epithelium in culture. We hypothesized that we could take advantage of the integral cochlear structure to maintain the overall inner ear architecture and improve sensory epithelium survival in culture. We showed that by culturing adult mouse cochlea with the (surrounding) bone intact, the supporting cells (SCs) survived and almost all hair cells (HCs) degenerated. To evaluate the utility of the explant culture system, we demonstrated that the overexpression of Atoh1, an HC fate-determining factor, is sufficient to induce transdifferentiation of adult SCs to HC-like cells (HCLCs). Transdifferentiation-derived HCLCs resemble developmentally young HCs and are able to attract adult ganglion neurites. Furthermore, using a damage model, we showed that degenerated adult ganglions respond to regenerated HCLCs by directional neurite outgrowth that leads to HCLC-neuron contacts, strongly supporting the intrinsic properties of the HCLCs in establishing HCLC-neuron connections. The adult whole cochlear explant culture is suitable for diverse studies of the adult inner ear including regeneration, HC-neuron pathways, and inner ear drug screening.

Development of Ultrasensitive Biomimetic Auditory Hair Cells Based on Piezoresistive Hydrogel Nanocomposites.

With an ageing population, hearing disorders are predicted to rise considerably in the following decades. Thus, developing a new class of artificial auditory system has been highlighted as one of the most exciting research topics for biomedical applications. Herein, a design of a biocompatible piezoresistive-based artificial hair cell sensor is presented consisting of a highly flexible and conductive polyvinyl alcohol (PVA) nanocomposite with vertical graphene nanosheets (VGNs). The bilayer hydrogel sensor demonstrates excellent performance to mimic biological hair cells, responding to acoustic stimuli in the audible range between 60 Hz to 20 kHz. The sensor output demonstrates stable mid-frequency regions (∼4-9 kHz), with the greatest sensitivity as high frequencies (∼13-20 kHz). This is somewhat akin to the mammalian auditory system, which has remarkable sensitivity and sharp tuning at high frequencies due to the "active process". This work validates the PVA/VGN sensor as a potential candidate to play a similar functional role to that of the cochlear hair cells, which also operate over a wide frequency domain in a viscous environment. Further characterizations of the sensor show that increasing the sound amplitude results in higher responses from the sensor while taking it to the depth drops the sensor outputs due to attenuation of sound in water. Meanwhile, the acoustic pressure distribution of sound waves is predicted through finite element analysis, whereby the numerical results are in perfect agreement with experimental data. This proof-of-concept work creates a platform for the future design of susceptible, flexible biomimetic sensors to closely mimic the biological cochlea.

The Kinocilia of Cochlear Hair Cells: Structures, Functions, and Diseases.

Primary cilia are evolutionarily conserved and highly specialized organelles that protrude from cell membranes. Mutations in genes encoding ciliary proteins can cause structural and functional ciliary defects and consequently multiple diseases, collectively termed ciliopathies. The mammalian auditory system is responsible for perceiving external sound stimuli that are ultimately processed in the brain through a series of physical and biochemical reactions. Here we review the structure and function of the specialized primary cilia of hair cells, termed kinocilia, found in the mammalian auditory system. We also discuss areas that might prove amenable for therapeutic management of auditory ciliopathies.

Visual Signals in the Mammalian Auditory System.

Coordination between different sensory systems is a necessary element of sensory processing. Where and how signals from different sense organs converge onto common neural circuitry have become topics of increasing interest in recent years. In this article, we focus specifically on visual-auditory interactions in areas of the mammalian brain that are commonly considered to be auditory in function. The auditory cortex and inferior colliculus are two key points of entry where visual signals reach the auditory pathway, and both contain visual- and/or eye movement-related signals in humans and other animals. The visual signals observed in these auditory structures reflect a mixture of visual modulation of auditory-evoked activity and visually driven responses that are selective for stimulus location or features. These key response attributes also appear in the classic visual pathway but may play a different role in the auditory pathway: to modify auditory rather than visual perception. Finally, while this review focuses on two particular areas of the auditory pathway where this question has been studied, robust descending as well as ascending connections within this pathway suggest that undiscovered visual signals may be present at other stages as well.

Development in the Mammalian Auditory System Depends on Transcription Factors

We review the molecular basis of several transcription factors (Eya1, Sox2), including the three related genes coding basic helix–loop–helix (bHLH; see abbreviations) proteins (Neurog1, Neurod1, Atoh1) during the development of spiral ganglia, cochlear nuclei, and cochlear hair cells. Neuronal development requires Neurog1, followed by its downstream target Neurod1, to cross-regulate Atoh1 expression. In contrast, hair cells and cochlear nuclei critically depend on Atoh1 and require Neurod1 expression for interactions with Atoh1. Upregulation of Atoh1 following Neurod1 loss changes some vestibular neurons’ fate into “hair cells”, highlighting the significant interplay between the bHLH genes. Further work showed that replacing Atoh1 by Neurog1 rescues some hair cells from complete absence observed in Atoh1 null mutants, suggesting that bHLH genes can partially replace one another. The inhibition of Atoh1 by Neurod1 is essential for proper neuronal cell fate, and in the absence of Neurod1, Atoh1 is upregulated, resulting in the formation of “intraganglionic” HCs. Additional genes, such as Eya1/Six1, Sox2, Pax2, Gata3, Fgfr2b, Foxg1, and Lmx1a/b, play a role in the auditory system. Finally, both Lmx1a and Lmx1b genes are essential for the cochlear organ of Corti, spiral ganglion neuron, and cochlear nuclei formation. We integrate the mammalian auditory system development to provide comprehensive insights beyond the limited perception driven by singular investigations of cochlear neurons, cochlear hair cells, and cochlear nuclei. A detailed analysis of gene expression is needed to understand better how upstream regulators facilitate gene interactions and mammalian auditory system development.

Mathematical model of the auditory nerve response to stimulation by a micro-machined cochlea.

We report a novel mathematical model of an artificial auditory system consisting of a micro-machined cochlea and the auditory nerve response it evokes. The modeled micro-machined cochlea is one previously realized experimentally by mimicking functions of the cochlea [Shintaku et al, Sens. Actuat. 158 (2010) 183-192; Inaoka et al, Proc. Natl. Acad. Sci. USA 108 (2011) 18390-18395]. First, from the viewpoint of mechanical engineering, the frequency characteristics of a model device were experimentally investigated to develop an artificial basilar membrane based on a spring-mass-damper system. In addition, a nonlinear feedback controller mimicking the function of the outer hair cells was incorporated in this experimental system. That is, the developed device reproduces the proportional relationship between the oscillation amplitude of the basilar membrane and the cube root of the sound pressure observed in the mammalian auditory system, which is what enables it to have a wide dynamic range, and the characteristics of the control performance were evaluated numerically and experimentally. Furthermore, the stimulation of the auditory nerve by the micro-machined cochlea was investigated using the present mathematical model, and the simulation results were compared with our previous experimental results from animal testing [Shintaku et al, J. Biomech. Sci. Eng. 8 (2013) 198-208]. The simulation results were found to be in reasonably good agreement with those from the previous animal test; namely, there exists a threshold at which the excitation of the nerve starts and a saturation value for the firing rate under a large input. The proposed numerical model was able to qualitatively reproduce the results of the animal test with the micro-machined cochlea and is thus expected to guide the evaluation of micro-machined cochleae for future animal experiments.

High frequency sensitivity to interaural onset time differences in the bat inferior colliculus

Many neurons in the auditory midbrain are tuned to binaural cues. Two prominent binaural cues are the interaural level difference (ILD) and the interaural time difference (ITD). The ITD cue can further be subdivided into the ongoing envelope ITD cues and transient onset ITD cues. More is known about the sensitivity of single neurons to ongoing envelope ITDs compared to transient onset ITDs in the mammalian auditory system, particularly in bats. The current study examines the response properties of single neurons in the inferior colliculus (IC) of the big brown bat, Eptesicus fuscus, to onset ITDs in response to high frequency pure tones. Measures of neurons’ dynamic ITD response revealed an average change of 36% of its maximum response within the behaviorally relevant range of ITDs (±50 µs). Across all IC neurons, we measured an average time-intensity trading ratio of 30 µs/dB in the sensitivity of the ITD response function to changing ILDs. Minimum and maximum ITD responses were clustered within a narrow range of ITDs. The average peak in the ITD response function was at 268 µs, a finding that is consistent with other non-echolocating mammals. Some ITD-sensitive neurons also showed weak facilitation of maximum response during binaural stimulation, compared to monaural stimulation. These results suggest that echolocating bats possess the potential to use onset ITD cues to assist in the azimuthal sound localization of ultrasonic frequencies.

Lmx1a and Lmx1b are Redundantly Required for the Development of Multiple Components of the Mammalian Auditory System

The inner ear, projections, and brainstem nuclei are essential components of the auditory and vestibular systems. It is believed that the evolution of complex systems depends on duplicated sets of genes. The contribution of duplicated genes to auditory or vestibular system development, however, is poorly understood. We describe that Lmx1a and Lmx1b, which originate from the invertebrate Lmx1b-like gene, redundantly regulate development of multiple essential components of the mammalian auditory/vestibular systems. Combined, but not individual, loss of Lmx1a/b eliminated the auditory inner ear organ of Corti (OC) and disrupted the spiral ganglion, which was preceded by a diminished expression of their critical regulator Pax2. Innervation of the remaining inner ear vestibular organs revealed unusual sizes or shapes and was more affected compared to Lmx1a/b single-gene mutants. Individual loss of Lmx1a/b genes did not disrupt brainstem auditory nuclei or inner ear central projections. Combined loss of Lmx1a/b, however, eliminated excitatory neurons in cochlear/vestibular nuclei, and also the expression of a master regulator Atoh1 in their progenitors in the lower rhombic lip (RL). Finally, in Lmx1a/b double mutants, vestibular afferents aberrantly projected to the roof plate. This phenotype was associated with altered expression of Wnt3a, a secreted ligand of the Wnt pathway that regulates pathfinding of inner ear projections. Thus, Lmx1a/b are redundantly required for the development of the mammalian inner ear, inner ear central projections, and cochlear/vestibular nuclei.