Mammalian Ear Research Articles

Convergent evolution in Afrotheria and non-afrotherians demonstrates high evolvability of the mammalian inner ear

Evolutionary convergence in distantly related species is among the most convincing evidence of adaptive evolution. The mammalian ear, responsible for balance and hearing, is not only characterised by its spectacular evolutionary incorporation of several bones of the jaw, it also varies considerably in shape across modern mammals. Using a multivariate approach, we show that in Afrotheria, a monophyletic clade with morphologically and ecologically highly disparate species, inner ear shape has evolved similar adaptations as in non-afrotherian mammals. We identify four eco-morphological trait combinations that underlie this convergence. The high evolvability of the mammalian ear is surprising: Nowhere else in the skeleton are different functional units so close together; it includes the smallest bones of the skeleton, encapsulated within the densest bone. We suggest that this evolvability is a direct consequence of the increased genetic and developmental complexity of the mammalian ear compared to other vertebrates.

Transdifferentiation is temporally uncoupled from progenitor pool expansion during hair cell regeneration in the zebrafish inner ear.

Death of mechanosensory hair cells in the inner ear is a common cause of auditory and vestibular impairment in mammals, which have a limited ability to regrow these cells after damage. In contrast, non-mammalian vertebrates, including zebrafish, can robustly regenerate hair cells after severe organ damage. The zebrafish inner ear provides an understudied model system for understanding hair cell regeneration in organs that are highly conserved with their mammalian counterparts. Here, we quantitatively examine hair cell addition during growth and regeneration of the larval zebrafish inner ear. We used a genetically encoded ablation method to induce hair cell death and we observed gradual regeneration with correct spatial patterning over a 2-week period following ablation. Supporting cells, which surround and are a source of new hair cells, divide in response to hair cell ablation, expanding the possible progenitor pool. In parallel, nascent hair cells arise from direct transdifferentiation of progenitor pool cells temporally uncoupled from supporting cell division. These findings reveal a previously unrecognized mechanism of hair cell regeneration with implications for how hair cells may be encouraged to regenerate in the mammalian ear.

Bioinspired Fano-like resonant transmission: frequency selective impedance matching

The study of the impedance mismatch between the device and its surroundings is crucial when building an acoustic device to obtain optimal performance. In reality, a high impedance mismatch would prohibit energy from being transmitted over the interface, limiting the amount of energy that the device could treat. In general, this is solved by using acoustic impedance matching layers, such as gradients, similar to what is done in optical coatings. The simplest form of such a gradient can be considered as an intermediate layer with certain qualities resting between the two media to impedance match, and requiring a minimum thickness of at least one quarter wavelength of the lowest frequency under consideration. The desired combination(s) of the (limited) available elastic characteristics and densities has traditionally determined material selection. Nature, which is likewise limited by the use of a limited number of materials in the construction of biological structures, demonstrates a distinct approach in which the design space is swept by modifying certain geometrical and/or material parameters. The middle ear of mammals and the lateral line of fishes are both instances of this method, with the latter already incorporating an architecture of distributed impedance matched underwater layers. In this paper, we develop a resonant mechanism whose properties can be modified to give impedance matching at different frequencies by adjusting a small set of geometrical parameters. The mechanism in question, like the lateral line organ, is intended to serve as the foundation for the creation of an impedance matching meta-surface. A computational study and parameter optimization show that it can match the impedance of water and air in a deeply sub-wavelength zone.

EpCAM regulates hair cell development and regeneration in the zebrafish lateral line.

The zebrafish lateral line mechanosensory system shares considerable morphological and molecular similarities with the inner ear. In particular, mechanosensory hair cells are responsible for transducing sensory stimuli in both structures. The epithelia cell adhesion molecule (EpCAM) is expressed in the cells of the inner ear of mammals and in the lateral lines system of fish. EpCAM regulates the many cellular functions including adhesion, migration, proliferation, and differentiation. In this study, we use the epcam jh79 mutant zebrafish line to determine that EpCAM function is required for proper development and regeneration of posterior lateral line hair cells.

An Eocene insect could hear conspecific ultrasounds and bat echolocation

Hearing has evolved independently many times in the animal kingdom and is prominent in various insects and vertebrates for conspecific communication and predator detection. Among insects, katydid (Orthoptera: Tettigoniidae) ears are unique, as they have evolved outer, middle, and inner ear components, analogous in their biophysical principles to the mammalian ear. The katydid ear consists of two paired tympana located in each foreleg. These tympana receive sound externally on the tympanum surface (usually via pinnae) or internally via an ear canal (EC). The EC functions to capture conspecific calls and low frequencies, while the pinnae passively amplify higher-frequency ultrasounds including bat echolocation. Together, these outer ear components provide enhanced hearing sensitivity across a dynamic range of over 100 kHz. However, despite a growing understanding of the biophysics and function of the katydid ear, its precise emergence and evolutionary history remains elusive. Here, using microcomputed tomography (μCT) scanning, we recovered geometries of the outer ear components and wings of an exceptionally well-preserved katydid fossilized in Baltic amber (∼44 million years [Ma]). Using numerical and theoretical modeling of the wings, we show that this species was communicating at a peak frequency of 31.62 (± 2.27) kHz, and we demonstrate that the ear was biophysically tuned to this signal and to providing hearing at higher-frequency ultrasounds (>80 kHz), likely for enhanced predator detection. The results indicate that the evolution of the unique ear of the katydid, with its broadband ultrasonic sensitivity and analogous biophysical properties to the ears of mammals, emerged in the Eocene.

Elevator-like movements of prestin mediate outer hair cell electromotility

The outstanding acuity of the mammalian ear relies on cochlear amplification, an active mechanism based on the electromotility (eM) of outer hair cells. eM is a piezoelectric mechanism generated by little-understood, voltage-induced conformational changes of the anion transporter homolog prestin (SLC26A5). We used a combination of molecular dynamics (MD) simulations and biophysical approaches to identify the structural dynamics of prestin that mediate eM. MD simulations showed that prestin samples a vast conformational landscape with expanded (ES) and compact (CS) states beyond previously reported prestin structures. Transition from CS to ES is dominated by the translational-rotational movement of prestin’s transport domain, akin to elevator-type substrate translocation by related solute carriers. Reversible transition between CS and ES states was supported experimentally by cysteine accessibility scanning, cysteine cross-linking between transport and scaffold domains, and voltage-clamp fluorometry (VCF). Our data demonstrate that prestin’s piezoelectric dynamics recapitulate essential steps of a structurally conserved ion transport cycle.

Efferent axons in the zebrafish lateral line degenerate following sensory hair cell ablation

The zebrafish lateral line is a frequently used model to study the mechanisms behind peripheral neuronal innervation of sensory organs and the regeneration thereof. The lateral line system consists of neuromasts, a cluster of protruding hair cells, which are innervated by sensory afferent and modulatory efferent neurons. These flow-sensing hair cells are similar to the hair cells in the mammalian ear. Though, while hair cell loss in humans is irreversible, the zebrafish neuromasts are regarded as the fastest regenerating structure in vertebrates, making them an ideal model to study regeneration. However, one component of the lateral line system, the efferent projections, has largely been omitted in regenerative studies. Here, for the first time, we bring insights into the fate of efferent axons during ablation and regeneration of the hair cells in the zebrafish lateral line. Our behavioral analysis showed functional recovery of hair cells and sensory transmission within 48 h and their regeneration were in line with previous studies. Analysis of the inhibitory efferent projections revealed that in approximately half the cases the inhibitory efferent axons degenerated, which was never observed for the sensory afferent axons. Quantification of hair cells following ablation suggests that the presence of mature hair cells in the neuromast may prevent axon degeneration. Within 120 h, degenerated efferent axons regenerated along the axonal tract of the lateral line. Reanalysis of published single cell neuromast data hinted to a role for Bdnf in the survival of efferent axons. However, sequestering Bdnf, blocking the Trk-receptors, and inhibiting the downstream ERK-signaling, did not induce axon degeneration, indicating that efferent survival is not mediated through neurotrophic factors. To further explore the relation between hair cells and efferent projections, we generated atoh1a mutants, where mature hair cells never form. In larvae lacking hair cells, inhibitory efferent projections were still present, following the tract of the sensory afferent without displaying any innervation. Our study reveal the fate of efferent innervation following hair cell ablation and provide insights into the inherent differences in regeneration between neurons in the peripheral and central nervous system.

Cross-species analysis and comparison of the inner ear between chickens and mice.

The inner ear of mammals includes the cochlea and vestibule, which house specialized hair cells that are responsible for hearing and balance, respectively. While cochlear hair cells fail to regenerate following damage, those of the utricle, which is part of the vestibular apparatus, show partial regeneration. In birds, the macula lagena, a unique ear structure in this clade, has the ability to regenerate hair cells similarly to the utricle. Many studies have sought to explain regeneration in terms of evolution and species differences. However, it remains unclear what the cellular and molecular basis is behind the differences in inner ear structures and between avians and mammals. In the present study, we first investigated the anatomical structures of the inner ear of both chickens and rodents. We then performed RNA sequencing (RNA-Seq) and made cross-species analyses of the expression of homologous genes obtained from the inner ear tissue from both chickens and mice. Finally, we focused on the lagena, the basilar papilla, and the utricle in chickens and identified differentially expressed genes between tissues and determined the expression patterns of genes involved in inner ear structure formation by single-cell RNA sequencing and bulk RNA-Seq. We concluded that the cellular and molecular composition of the lagena is more similar to that of the utricle than the cochlea. Taken together, our study provides a valuable resource for the study of inner ear evolution and development.

ESPRESS.0: Eustachian Tube-Inspired Tactile Sensor Exploiting Pneumatics for Range Extension and SenSitivity Tuning.

Optimising the sensitivity of a tactile sensor to a specific range of stimuli magnitude usually compromises the sensor's widespread usage. This paper presents a novel soft tactile sensor capable of dynamically tuning its stiffness for enhanced sensitivity across a range of applied forces, taking inspiration from the Eustachian tube in the mammalian ear. The sensor exploits an adjustable pneumatic back pressure to control the effective stiffness of its 20 mm diameter elastomer interface. An internally translocated fluid is coupled to the membrane and optically tracked to measure physical interactions at the interface. The sensor can be actuated by pneumatic pressure to dynamically adjust its stiffness. It is demonstrated to detect forces as small as 0.012 N, and to be sensitive to a difference of 0.006 N in the force range of 35 to 40 N. The sensor is demonstrated to be capable of detecting tactile cues on the surface of objects in the sub-millimetre scale. It is able to adapt its compliance to increase its ability for distinguishing between stimuli with similar stiffnesses (0.181 N/mm difference) over a large range (0.1 to 1.1 N/mm) from only a 0.6 mm deep palpation. The sensor is intended to interact comfortably with skin, and the feasibility of its use in palpating tissue in search of hard inclusions is demonstrated by locating and estimating the size of a synthetic hard node embedded 20 mm deep in a soft silicone sample. The results suggest that the sensor is a good candidate for tactile tasks involving unpredictable or unknown stimuli.

A novel cis-regulatory element drives early expression of Nkx3.2 in the gnathostome primary jaw joint.

The acquisition of movable jaws was a major event during vertebrate evolution. The role of NK3 homeobox 2 (Nkx3.2) transcription factor in patterning the primary jaw joint of gnathostomes (jawed vertebrates) is well known, however knowledge about its regulatory mechanism is lacking. In this study, we report a proximal enhancer element of Nkx3.2 that is deeply conserved in most gnathostomes but undetectable in the jawless hagfish and lamprey. This enhancer is active in the developing jaw joint region of the zebrafish Danio rerio, and was thus designated as jaw joint regulatory sequence 1 (JRS1). We further show that JRS1 enhancer sequences from a range of gnathostome species, including a chondrichthyan and mammals, have the same activity in the jaw joint as the native zebrafish enhancer, indicating a high degree of functional conservation despite the divergence of cartilaginous and bony fish lineages or the transition of the primary jaw joint into the middle ear of mammals. Finally, we show that deletion of JRS1 from the zebrafish genome using CRISPR/Cas9 results in a significant reduction of early gene expression of nkx3.2 and leads to a transient jaw joint deformation and partial fusion. Emergence of this Nkx3.2 enhancer in early gnathostomes may have contributed to the origin and shaping of the articulating surfaces of vertebrate jaws.

Overturning the mechanisms of cochlear amplification via area deformations of the organ of Corti.

The mammalian ear embeds a cellular amplifier that boosts sound-induced hydromechanical waves as they propagate along the cochlea. The operation of this amplifier is not fully understood and is difficult to disentangle experimentally. In the prevailing view, cochlear waves are amplified by the piezo-electric action of the outer hair cells (OHCs), whose cycle-by-cycle elongations and contractions inject power into the local motion of the basilar membrane (BM). Concomitant deformations of the opposing (or "top") side of the organ of Corti are assumed to play a minor role and are generally neglected. However, analysis of intracochlear motions obtained using optical coherence tomography calls this prevailing view into question. In particular, the analysis suggests that (i) the net local power transfer from the OHCs to the BM is either negative or highly inefficient; and (ii) vibration of the top side of the organ of Corti plays a primary role in traveling-wave amplification. A phenomenological model derived from these observations manifests realistic cochlear responses and suggests that amplification arises almost entirely from OHC-induced deformations of the top side of the organ of Corti. In effect, the model turns classic assumptions about spatial impedance relations and power-flow direction within the sensory epithelium upside down.

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.

Interplay between traveling wave propagation and amplification at the apex of the mouse cochlea

Sounds entering the mammalian ear produce waves that travel from the base to the apex of the cochlea. An electromechanical active process amplifies traveling wave motions and enables sound processing over a broad range of frequencies and intensities. The cochlear amplifier requires combining the global traveling wave with the local cellular processes that change along the length of the cochlea given the gradual changes in hair cell and supporting cell anatomy and physiology. Thus, we measured basilar membrane (BM) traveling waves in vivo along the apical turn of the mouse cochlea using volumetric optical coherence tomography and vibrometry. We found that there was a gradual reduction in key features of the active process toward the apex. For example, the gain decreased from 23 to 19 dB and tuning sharpness decreased from 2.5 to 1.4. Furthermore, we measured the frequency and intensity dependence of traveling wave properties. The phase velocity was larger than the group velocity, and both quantities gradually decrease from the base to the apex denoting a strong dispersion characteristic near the helicotrema. Moreover, we found that the spatial wavelength along the BM was highly level dependent in vivo, such that increasing the sound intensity from 30 to 90 dB sound pressure level increased the wavelength from 504 to 874 μm, a factor of 1.73. We hypothesize that this wavelength variation with sound intensity gives rise to an increase of the fluid-loaded mass on the BM and tunes its local resonance frequency. Together, these data demonstrate a strong interplay between the traveling wave propagation and amplification along the length of the cochlea.

OD PĘCZKA KOMÓREK SŁUCHOWYCH DO BŁONY BĘBENKOWEJ

BackgroundAn earlier oscillator model for the generation of spontaneous otoacoustic emissions (SOAEs) from the lizard ear is extended with a connection of the oscillators to the basilar papilla, to make it possible that these SOAEs can be transported to the tympanic membrane, to be emitted.Material and methodsThe generators of spontaneous otoacoustic emissions are modelled as a one-dimensional array of Hopf-resonators. The resonators (or oscillators) are coupled to their neighbours, and to the basilar papilla. The papilla is modelled as a rigid structure, that is flexibly connected to its surroundings.ResultsFrequency spectra are given for different sets of coupling parameters, both for nearest neighbour coupling of the oscillators, and for coupling to the papilla, and also after the introduction of irregularities in the damping of the oscillators. Waterfall and density plots show clustering of the oscillators in frequency plateaus, and entrainment of a cluster of oscillators by an externally applied sinusoidal force. All these model outcomes correspond with characteristics of SOAEs emitted by real lizard ears.ConclusionsThe present model is a useful extension of an earlier model. Because its characteristics differ from that of a model that is used to describe the generation of SOAEs by mammalian ears, it revives the discussion whether different models are needed for SOAE generation in different animal species.

Size and shape of the semicircular canal of the inner ear: A new marker of pig domestication?

Domestication has led to many changes in domestic animal biology, including their anatomy. The shape of the inner ear, part of the mammalian ear, has been found particularly relevant for discriminating domesticated species, their hybrids or differentiating the wild and domestic populations of a single species. Here we assessed the use of the size and shape of the semicircular canals (SCC) of the inner ear as a marker of pig domestication. We studied a total of 63 petrosal bones belonging to wild boar (Sus scrofa, two populations) and domestic pigs (extensively and intensively reared specimens) that were µCT-scanned and from which the size and the shape of the inner ear were quantified through geometric morphometrics, analyzing the 3D coordinates of 6 landmarks and 60 sliding semilandmarks localized on the SCC and the common crus. The domestic pigs have larger SCC than the wild boar from which they also strongly differ in shape (correct cross validation of 95.5%, confidence interval: 92.3%-98.1%). Strong shape differences were detected between the two populations of wild boar, as well as a sexual size dimorphism. All together the results highlight the taxonomic discriminant power of the SCC of the inner ear shape, and its relevance for domestication studies.

Whistling While it Works: Spontaneous Otoacoustic Emissions and the Cochlear Amplifier.

Perhaps the most striking evidence for active processes operating within the inner ears of mammals and non-mammals alike is their ability to spontaneously produce sound. Predicted by Thomas Gold in 1948, some 30 years prior to their discovery, the narrow-band sounds now known as spontaneous otoacoustic emissions (SOAEs) remain incompletely understood, their origins controversial. Without a single equation in the main text, we review the essential concepts underlying the "local-" and "global-oscillator" frameworks for understanding SOAE generation. Comparing their key assumptions and predictions, we relate the two frameworks to unresolved questions about the biophysical mechanisms of cochlear amplification.

Mechanical overstimulation causes acute injury and synapse loss followed by fast recovery in lateral-line neuromasts of larval zebrafish

Excess noise damages sensory hair cells, resulting in loss of synaptic connections with auditory nerves and, in some cases, hair-cell death. The cellular mechanisms underlying mechanically induced hair-cell damage and subsequent repair are not completely understood. Hair cells in neuromasts of larval zebrafish are structurally and functionally comparable to mammalian hair cells but undergo robust regeneration following ototoxic damage. We therefore developed a model for mechanically induced hair-cell damage in this highly tractable system. Free swimming larvae exposed to strong water wave stimulus for 2 hr displayed mechanical injury to neuromasts, including afferent neurite retraction, damaged hair bundles, and reduced mechanotransduction. Synapse loss was observed in apparently intact exposed neuromasts, and this loss was exacerbated by inhibiting glutamate uptake. Mechanical damage also elicited an inflammatory response and macrophage recruitment. Remarkably, neuromast hair-cell morphology and mechanotransduction recovered within hours following exposure, suggesting severely damaged neuromasts undergo repair. Our results indicate functional changes and synapse loss in mechanically damaged lateral-line neuromasts that share key features of damage observed in noise-exposed mammalian ear. Yet, unlike the mammalian ear, mechanical damage to neuromasts is rapidly reversible.

A narrow ear canal reduces sound velocity to create additional acoustic inputs in a microscale insect ear

Located in the forelegs, katydid ears are unique among arthropods in having outer, middle, and inner components, analogous to the mammalian ear. Unlike mammals, sound is received externally via two tympanic membranes in each ear and internally via a narrow ear canal (EC) derived from the respiratory tracheal system. Inside the EC, sound travels slower than in free air, causing temporal and pressure differences between external and internal inputs. The delay was suspected to arise as a consequence of the narrowing EC geometry. If true, a reduction in sound velocity should persist independently of the gas composition in the EC (e.g., air, [Formula: see text]). Integrating laser Doppler vibrometry, microcomputed tomography, and numerical analysis on precise three-dimensional geometries of each experimental animal EC, we demonstrate that the narrowing radius of the EC is the main factor reducing sound velocity. Both experimental and numerical data also show that sound velocity is reduced further when excess [Formula: see text] fills the EC. Likewise, the EC bifurcates at the tympanal level (one branch for each tympanic membrane), creating two additional narrow internal sound paths and imposing different sound velocities for each tympanic membrane. Therefore, external and internal inputs total to four sound paths for each ear (only one for the human ear). Research paths and implication of findings in avian directional hearing are discussed.

Anatomy and Development of the Mammalian External Auditory Canal: Implications for Understanding Canal Disease and Deformity.

The mammalian ear is made up of three parts (the outer, middle, and inner ear), which work together to transmit sound waves into neuronal signals perceived by our auditory cortex as sound. This review focuses on the often-neglected outer ear, specifically the external auditory meatus (EAM), or ear canal. Within our complex hearing pathway, the ear canal is responsible for funneling sound waves toward the tympanic membrane (ear drum) and into the middle ear, and as such is a physical link between the tympanic membrane and the outside world. Unique anatomical adaptations, such as its migrating epithelium and cerumen glands, equip the ear canal for its function as both a conduit and a cul-de-sac. Defects in development, or later blockages in the canal, lead to congenital or acquired conductive hearing loss. Recent studies have built on decades-old knowledge of ear canal development and suggest a novel multi-stage, complex and integrated system of development, helping to explain the mechanisms underlying congenital canal atresia and stenosis. Here we review our current understanding of ear canal development; how this biological lumen is made; what determines its location; and how its structure is maintained throughout life. Together this knowledge allows clinical questions to be approached from a developmental biology perspective.

Kcnb1 plays a role in development of the inner ear

The function of the inner ear depends on the maintenance of high concentrations of K+ ions. The slow-inactivating delayed rectifier Kv2.1/KCNB1 channel works in the inner ear in mammals. The kcnb1 gene is expressed in the otic vesicle of developing zebrafish, suggesting its role in development of the inner ear. In the present study, we found that a Kcnb1 loss-of-function mutation affected development of the inner ear at multiple levels, including otic vesicle expansion, otolith formation, and the proliferation and differentiation of mechanosensory cells. This resulted in defects of kinocilia and stereocilia and abnormal function of the inner ear detected by behavioral assays. The quantitative transcriptional analysis of 75 genes demonstrated that the kcnb1 mutation affected the transcription of genes that are involved in K+ metabolism, cell proliferation, cilia development, and intracellular protein trafficking. These results demonstrate a role for Kv2.1/Kcnb1 channels in development of the inner ear in zebrafish.