Mechanics of the inner ear of the bullfrog (Rana catesbeiana): the contact membranes and the periotic canal

The frog inner ear consists of a complex of fluid-filled membranous sacs and canals containing eight distinct clusters of sensory hair cells. In this study we attempt to delineate the potential pathways for acoustic energy flow toward two of these clusters located within the amphibian papilla and the basilar papilla. Detailed morphological measurements of the periotic canal based on internal casts of the inner ear in the bullfrog (Rana catesbeiana) revealed that it is divided into a wide, tapered section and a narrower section comprised of two branches – one short and blind projecting into the endolymphatic space and another longer, terminating in the round window. Additionally, we used laser Doppler velocimetry to record the velocity responses of the contact membranes of the amphibian papilla and basilar papilla. We found that the acoustic energy flow through these two structures is frequency dependent such that the amphibian papilla contact membrane displays a peak velocity amplitude at frequencies less than 500 Hz, whereas the basilar papilla contact membrane velocity response exhibits a maximum above 1100 Hz. Our data advocate a mechanical substrate underlying the frequency segregation in the auditory nerve fibers innervating the amphibian papilla and the basilar papilla. Accepted: 9 March 2000     

Does the morphology of the ear of the Chinese bamboo rat (Rhizomys sinensis) show “Subterranean” characteristics?

In spite of the growing interest in rodents with subterranean activity in general and the spalacids (Spalacidae) in particular, little is known about the biology of most members of this clade, such as the Chinese bamboo rat (Rhizomys sinensis). Here, we analyzed the ear morphology of R. sinensis with respect to hearing specialization for subterranean or aboveground modes of communication. It is well‐known that ecology and style of life of a particular species can be reflected in morphology of its ear, its hearing and vocalization, so we expect that such information could provide us insight into its style of life and its sensory environment. The ratio between the eardrum and stapedial footplate areas, which influences the efficiency of middle ear sound transmission, suggests low hearing sensitivity, as is typical for subterranean species. The cochlea had 3.25 coils and resembled species with good low frequency hearing typical for subterranean mammals. The length of the basilar membrane was 18.9 ± 0.8 mm and its width slowly increased towards the cochlear apex from 60 to 85 μm. The mean density of outer hair cells was 344 ± 22 and of inner hair cells 114 ± 7.3 per 1 mm length of the organ of Corti, and increased apically. These values (except for relatively low hair cell density) usually characterize ears specialized for low frequency hearing. There was no evidence for an acoustic fovea. Apart of low hair cell density which is common in aboveground animals, this species has also relatively large auricles, suggesting the importance of sound localization during surface activity. The ear of the Chinese bamboo rat thus contains features typical for both aboveground and subterranean mammals and suggests that this spalacid has fossorial habits combined with regular aboveground activity. J. Morphol. 277:575–584, 2016. © 2016 Wiley Periodicals, Inc.     

An auditory fovea in the barn owl cochlea

The distribution of frequencies along the basilar papilla of the barn owl (Tyto alba) was studied by labelling small groups of primary auditory neurones of defined frequency response and tracing them to their peripheral innervation sites. The exact location of marked neurones was determined in cochlear wholemounts with the aid of a special surface preparation technique. The average basilar papilla length (in fixed, embedded specimens) was 10.74 mm.The resulting frequency map shows the basic vertebrate pattern with the lowest frequencies represented apically and increasingly higher frequencies mapped at progressively more basal locations. However, the length of basilar papilla devoted to different frequency ranges, i.e. the space per octave, varies dramatically in the barn owl. The lower frequencies (up to 2 kHz) show values between about 0.35 and 1 mm/octave, which are roughly equivalent to values reported for other birds. Above that, the space increases enormously, the highest octave (5–10 kHz) covering about 6 mm, or more than half of the length of the basilar papilla.Such an overrepresentation of a narrow, behaviourally very important frequency band is also seen in some bats, where it has been termed an acoustic or auditory fovea.Abbreviations CF characteristic frequency - HRP horseradish peroxidase - NA Nucleus angularis - NM Nucleus magnocellularis     

内耳减压病豚鼠听力和耳蜗组织的病理变化

研究探讨了内耳减压病豚鼠皮层听觉诱发电位阈值、耳蜗火棉胶切片、酶组织化学和透射电镜观察的变化。结果表明,豚鼠内耳减压病导致听力损失,耳蜗广泛的病理损害.毛细胞琥珀酸脱氢酶活性降低。提出了加压治疗内耳减压病时配合改善微循环、增加能量供应等见解。     

What did Morganucodon hear?

The structure of the middle and inner ear of Morganucodon , one of the oldest known mammals, is reviewed and compared to the structure of the ears of extant mammals, reptiles and birds with known auditory capabilities. Specifically, allometric relationships between ear dimensions (basilar-membrane length, tympanic-membrane area and stapes-footplate area) and specific features of the audiogram are defined in extant ears. These relationships are then used to make several predictions of auditory function in Morganucodon. The results point out that the ear structures of Morganucodon–Art similar in dimensions to ear structures in both extant small mammals–with predominantly high-frequency (10 kHz) auditory capabilities, and reptiles and birds- with better low and middle-frequency hearing (< 5 kHz). Although the allometric analysis cannot by itself determine whether Morganucodon heard more like present-day small mammals, or birds and reptiles, the apparent stiffness of the Morganucodon middle ear is both more consistent with the high-frequency mammalian middle ear and would act to decrease the sensitivity of a bird-reptile middle ear to low-frequency sound. Several likely hearing scenarios for Morganucodon are defined, including a scenario in which these animals had ears like those of modern small mammals that are selectively sensitive to high-frequency sounds, and a second scenario in which the Morganucodon ear was moderately sensitive to sounds of a narrow middle-frequency range (5–7 kHz) and relatively insensitive to sounds of higher or lower frequency. The evidence needed to substantiate either scenario includes some objective measure of the stiffness of the Morganucodon ossicular system, while a key datum needed to distinguish between the two hypotheses includes confirmation of the presence or absence of a cochlear lamina in the Morganucodon inner ear.     

Tympanic and extratympanic sound transmission in the leopard frog

Summary The inner ear of the leopard frog,Rana pipiens, receives sound via two separate pathways: the tympanic-columellar pathway and an extratympanic route. The relative efficiency of the two pathways was investigated. Laser interferometry measurements of tympanic vibration induced by free-field acoustic stimulation reveal a broadly tuned response with maximal vibration at 800 and 1500 Hz. Vibrational amplitude falls off rapidly above and below these frequencies so that above 2 kHz and below 300 Hz tympanic vibration is severely reduced. Electrophysiological measurements of the thresholds of single eighth cranial nerve fibers from both the amphibian and basilar papillae in response to pure tones were made in such a way that the relative efficiency of tympanic and extratympanic transmission could be assessed for each fiber. Thresholds for the two routes are very similar up to 1.0 kHz, above which tympanic transmission eventually becomes more efficient by 15–20 dB. By varying the relative phase of the two modes of stimulation, a reduction of the eighth nerve response can be achieved. When considered together, the measurements of tympanic vibration and the measurements of tympanic and extratympanic transmission thresholds suggest that under normal conditions in this species (1) below 300 Hz extratympanic sound transmission is the main source of inner ear stimulation; (2) for most of the basilar papilla frequency range (i.e., above 1.2 kHz) tympanic transmission is more important; and (3) both routes contribute to the stimulation of amphibian papilla fibers tuned between those points. Thus acoustic excitation of the an uran's inner ear depends on a complex interac tion between tympanic and extratympanic sound transmission.Abbreviations dB SPL decibels sound pressure level re: 20 N/ m² - AP amphibian papilla - BP basilar papilla - BEF best excitatory frequency     

Mechanics of the exceptional anuran ear

The anuran ear is frequently used for studying fundamental properties of vertebrate auditory systems. This is due to its unique anatomical features, most prominently the lack of a basilar membrane and the presence of two dedicated acoustic end organs, the basilar papilla and the amphibian papilla. Our current anatomical and functional knowledge implies that three distinct regions can be identified within these two organs. The basilar papilla functions as a single auditory filter. The low-frequency portion of the amphibian papilla is an electrically tuned, tonotopically organized auditory end organ. The high-frequency portion of the amphibian papilla is mechanically tuned and tonotopically organized, and it emits spontaneous otoacoustic emissions. This high-frequency portion of the amphibian papilla shows a remarkable, functional resemblance to the mammalian cochlea.     

Distinct contributions from the hindbrain and mesenchyme to inner ear morphogenesis

A mature inner ear is a complex structure consisting of vestibular and auditory components. Microsurgical ablations, rotations, and translocations were performed in ovo to identify the tissues that control inner ear morphogenesis. We show that mesenchyme/ectoderm adjacent to the developing ear specifically governs the shape of vestibular components - the semicircular canals and ampullae - by conferring anteroposterior axial information to these structures. In contrast, removal of individual hindbrain rhombomeres adjacent to the developing ear preferentially affects the growth and morphogenesis of the auditory subdivision, the cochlear duct, or basilar papilla. Removal of rhombomere 5 affects cochlear duct growth, while rhombomere 6 removal affects cochlear growth and morphogenesis. Rotating rhombomeres 5 and 6 along the anteroposterior axis also impacts cochlear duct morphogenesis but has little effect on the vestibular components. Our studies indicate that discrete tissues, acting at a distance, control the morphogenesis of distinct elements of the inner ear. These results provide a basis for identifying factors that are essential to vestibular and auditory development in vertebrates.     

Development of hearing in vertebrates with special reference to anuran acoustic communication

Amphibians, specially anurans, are excellent model systems for studying acoustic communication. After hatching, anurans exist in two forms; these have two distinct mode of sound perception. Aquatic larvae are perceptive to waterborne sound stimuli; then, following metamorphosis, as terrestrial adults, perceptive to airborne sound stimuli. Added to this, the metamorphosing tadpole presents an equally interesting study as it could recapitulate the events which occurred during the evolution of hearing in vertebrates at the lime of the transition from aquatic to terrestrial life. Metamorphosis entails the loss of a prominent aquatic sensory system—the lateral line system—and the simultaneous gain of another, the inner ear, along with the coevolution of the tympanic middle ear, a basilar papilla and a periotic labyrinth in the inner ear. Another interesting feature is that anurans are believed to be the first terrestrial vertebrates to use vocalization as a part of their reproductive behaviour. Vocal communication plays an important role in behaviour, ranging from territorial defense to reproduction, and calls are classified according to the particular behaviors that they subserve. Adult male anurans produce a species-specific mating call which is used to attract conspecific females dung their mating season, and this call serves as a mechanism in maintaining reproductive isolation from other sympatric species.     

A model for energy flow in the inner ear of the bullfrog (Rana catesbeiana)

We present a quantitative mathematical model that represents the main features of the bullfrog inner ear. Calculated responses based on this model predict the observed frequency separation between the amphibian papilla and basilar papilla responses. The origin of this separation can be traced to the effect of the contact membranes on the impedance of the respective paths. Additionally, we calculated the input impedance of the periotic canal and showed that at low frequencies it acts as a bypass for most of the energy entering the ear, shunting it away from the amphibian-basilar papilla complex. As this shunting decreases with increasing frequency, we propose that the periotic canal functions as a protection mechanism to prevent overload of the amphibian papilla and basilar papilla during ventilation and for quasi-static pressure equalization. Our model explains the main features of the empirical data obtained from direct measurement of the amphibian papilla and basilar papilla contact membranes reported in an accompanying paper (this issue). Accepted: 9 March 2000     

Auditory processing in anurans.

Anurans (frogs and toads) represent an example of peripheral specialization of the auditory systems. Their inner ear contains two distinct auditory organs: the amphibian papilla and the basilar papilla. Each organ is tuned to different species-specific frequency ranges. Because of this peripheral specialization, anurans offer an excellent opportunity to explore neural decoding of complex sounds in the central auditory system.     

Double stimulation of the inner ear organs of an anuran species (Alytes cisternasii) with simple tonal advertisement calls

Midwife toads present one of the simplest calls in anurans, with the whole energy concentrated in a single band without frequency modulation. The tuning curves of the Iberian midwife toads Alytes cisternasii show the typical bimodal pattern in anurans, with two best excitatory frequencies at 0.412 kHz (corresponding to the amphibian papilla) and at 1.358 kHz (corresponding to the basilar papilla and matching the male call frequency). In this study, the hypothesis that complex calls arose in anurans because they were inherently more attractive to females, since they provided greater acoustic stimulation, was tested. However, our results indicate that splitting the call energy to stimulate both inner ear organs simultaneously, the male call is not more attractive to female midwife toads, but sometimes renders it unattractive. The biological role of the amphibian papilla is discussed in ecological and evolutionary terms.     

Acoustic,auditory, and morphological divergence in three species of neotropical frog

Advertisement calls, auditory tuning, and larynx and ear morphology were examined in 3 neotropical frogs, Hyla microcephala, H. phlebodes and H. ebraccata, H. microcephala has the highest call dominant frequency (6.068 kHz) and basilar papilla tuning (5.36 kHz). H. phlebodes and H. ebraccata calls have lower dominant frequencies (3.832 and 3.197 kHz respectively) and basilar papilla tuning (2.79 and 2.56 kHz). The primary call notes of H. ebraccata are longer (181.6 ms) than those of H. microcephala (95.5 ms) or H. phlebodes (87.3 ms). Morphometric analysis suggests that temporal call features differ as laryngeal musculature changes, in the process changing the overall size of the larynx. The spectral aspects of the call differ as head size, and hence the size of its resonating and radiating structures, changes, modifying the dominant frequency of calls by accentuating their higher harmonics when head size decreases. Decreasing head size decreases the size of the middle and inner ear chambers, changing the mechanical tuning of the ear in the same direction as the change in dominant frequency. These changes result in divergent spectral-temporal characteristics of both the sending and receiving portions of the acoustic communication system underlying social behavior in these frogs.Abbreviations AP amphibian papilla - BEF best excitatory frequency - BP basilar papilla - dB SPL decibels sound pressure level re:20 N/m²     

Extracellular matrices associated with the apical surfaces of sensory epithelia in the inner ear: molecular and structural diversity

The ultrastructure and molecular composition of the extracellular matrices that are associated with the apical surfaces of the mechanosensory epithelia in the mouse inner ear are compared. A progressive increase in molecular and structural organization is observed, with the cupula being the simplest, the otoconial membrane exhibiting an intermediate degree of complexity, and the tectorial membrane being the most elaborate of the three matrices. These differences may reflect changes that occurred in the acellular membranes of the inner ear as a mammalian hearing organ arose during evolution from a simple equilibrium receptor. A comparison of the molecular composition of the acellular membranes in the chick inner ear suggests the auditory epithelium and the striolar region of the maculae are homologous, indicating the basilar papilla may have evolved from the striolar region of an otolithic organ. A comparison of the tectorial membranes in the chick cochlear duct and the mouse cochlea reveals differences in the structure of the noncollagenous matrix in the two species that may result from differences in the stochiometry of alpha- and beta-tectorin and/or differences in the post-translational modification of alpha-tectorin. This comparison also indicates that the appearance of collagen in the mammalian tectorial membrane may have been a major step in the evolution of an electromechanically tuned vertebrate hearing organ that operates over an extended frequency range.     

FGFR3 expression during development and regeneration of the chick inner ear sensory epithelia.

Several studies suggest fibroblast growth factor receptor 3 (FGFR3) plays a role in the development of the auditory epithelium in mammals. We undertook a study of FGFR3 in the developing and mature chicken inner ear and during regeneration of this epithelium to determine whether FGFR3 shows a similar pattern of expression in birds. FGFR3 mRNA is highly expressed in most support cells in the mature chick basilar papilla but not in vestibular organs of the chick. The gene is expressed early in the development of the basilar papilla. Gentamicin treatment sufficient to destroy hair cells in the basilar papilla causes a rapid, transient downregulation of FGFR3 mRNA in the region of damage. In the initial stages of hair cell regeneration, the support cells that reenter the mitotic cycle in the basilar papilla do not express detectable levels of FGFR3 mRNA. However, once the hair cells have regenerated in this region, the levels of FGFR3 mRNA and protein expression rapidly return to approximate those in the undamaged epithelium. These results indicate that FGFR3 expression changes after drug-induced hair cell damage to the basilar papilla in an opposite way to that found in the mammalian cochlea and may be involved in regulating the proliferation of support cells.     

The cellular organization and nervous supply of the basilar papilla in the lizard,Calotes versicolor

Summary The basilar papilla of the lizard Calotes versicolor contains about 225 sensory cells. These are of two types: the short-haired type A cells in the ventral (apical) part of the organ, and the type B cells with long hair bundles, in the dorsal (basal) part of the organ. The type A cells are unidirectionally oriented and are covered by a tectorial membrane while the type B cells lack a covering structure and their hair bundles are oriented bidirectionally. Apart from those differences, the type A and type B cells are similar. They are columnar, and display the features common to most sensory cells in inner ear epithelia. The sensory cells are separated by supporting cells, which have long slender processes that keep the sensory cells apart. Close to the surface of the basilar papilla a terminal bar of specialized junctions interlocks adjacent cells. Below this, adjacent supporting cells are linked by an occluding junction.The cochlear nerve enters from the medial (neural) aspect. The fibres of the nerve lose their myelin sheaths as they enter the basilar papilla. Each sensory cell is associated with several nerve endings. All the nerves identified were afferent. Marked variations were seen between nerve endings in the basilar papilla, but no morphological equivalents of any functional differences were observed.This work is supported by grant no. B76-12X-00720-11A from the Swedish Medical Research Council, and by funds from the Karolinska Institute, Stockholm, Sweden.     

Scaling of the marsupial middle ear and its functional significance

The marsupial middle ear performs an anatomical impedance matching for acoustic energy travelling in air to reach the cochlea. The size of the middle ear sets constraints for the frequencies transmitted. For generalized placental mammals, it has been shown that the limit for high-frequency hearing can be predicted on the basis of middle ear ossicle mass, provided that the ears fulfil requirements of isometry. We studied the interspecific size variation of the middle ear in 23 marsupial species, with the following measurable parameters: skull mass, condylobasal length, ossicular masses for malleus, incus and stapes, tympanic membrane area, oval window area, and lever arm lengths for malleus and incus. Our results show that the middle ear size grows with negative allometry in relation to body size and that the internal proportions of the marsupial middle ear are largely isometric. This resembles the situation in placental mammals and allows us to use their isometric middle ear model to predict the high-frequency hearing limit for marsupials. We found that the isometry model predicts the high-frequency hearing limit for different marsupials well, indicating that marsupials can be used as auditory models for general therian mammalian hearing. At very high frequencies, other factors, such as the inner ear, seem to constrain mammalian hearing.     

Has the evolution of complexity in the amphibian papilla influenced anuran speciation rates?

For anurans, increasing complexity of the inner ear has been correlated with speciation rates. The evolution of a complex amphibian papilla (AP) is thought to have facilitated speciation by extending the range of frequencies over which mating calls may diverge. Although this example has been proposed to represent a key innovation, the mechanism by which the AP is thought to promote speciation makes the questionable assumption that anurans generally use the AP for detection of their mating calls. This study uses mating calls from 852 species to test this assumption. Surprisingly, the calls of most species are not detected by the AP but by a second organ, the basilar papilla (BP). This refutes the role of AP complexity in facilitating call divergence and hence, speciation. Future research into the evolution of acoustically mediated reproductive isolation should focus instead on the BP as it may play a more critical role in anuran speciation.     

Consequences of Location-Dependent Organ of Corti Micro-Mechanics

The cochlea performs frequency analysis and amplification of sounds. The graded stiffness of the basilar membrane along the cochlear length underlies the frequency-location relationship of the mammalian cochlea. The somatic motility of outer hair cell is central for cochlear amplification. Despite two to three orders of magnitude change in the basilar membrane stiffness, the force capacity of the outer hair cell’s somatic motility, is nearly invariant over the cochlear length. It is puzzling how actuators with a constant force capacity can operate under such a wide stiffness range. We hypothesize that the organ of Corti sets the mechanical conditions so that the outer hair cell’s somatic motility effectively interacts with the media of traveling waves—the basilar membrane and the tectorial membrane. To test this hypothesis, a computational model of the gerbil cochlea was developed that incorporates organ of Corti structural mechanics, cochlear fluid dynamics, and hair cell electro-physiology. The model simulations showed that the micro-mechanical responses of the organ of Corti are different along the cochlear length. For example, the top surface of the organ of Corti vibrated more than the bottom surface at the basal (high frequency) location, but the amplitude ratio was reversed at the apical (low frequency) location. Unlike the basilar membrane stiffness varying by a factor of 1700 along the cochlear length, the stiffness of the organ of Corti complex felt by the outer hair cell remained between 1.5 and 0.4 times the outer hair cell stiffness. The Y-shaped structure in the organ of Corti formed by outer hair cell, Deiters cell and its phalange was the primary determinant of the elastic reactance imposed on the outer hair cells. The stiffness and geometry of the Deiters cell and its phalange affected cochlear amplification differently depending on the location.