Hair Cells Of Cochlea Research Articles

The frequency limits to transduction in hair cells of the turtle cochlea

During transduction in auditory hair cells, displacements of the hair bundle modulate an inward transducer current; this then evokes a receptor potential that is filtered by the electrical properties of the cell membrane. The frequency characteristics of these steps have been studied by recording membrane currents in isolated turtle hair cells. When the hair bundle is rapidly deflected, the transducer current is activated with a maximum time constant of about 0.1 to 0.2 ms at 23 °C, thus faithfully encoding frequencies below 1 kHz. The ensuing receptor potentials are shaped by a sharply tuned resonance resulting from the interaction of a voltage-dependent Ca2+ current and a Ca2+-activated K+ current. The activation time constant for the K+ current varies inversely with, and largely determines, the resonant frequency of the hair cell in the range 20 to 600 Hz. The time constant of the Ca2+ current (0.3 ms) is rapid compared to the K+ current but must impose a corner frequency of about 500 Hz on transmission across the afferent synapse.

Synaptic bodies in the different rows of outer hair cells in the guinea pig cochlea.

The afferent synapses of the outer hair cells (OHCs) of the cochlea are peculiar, insofar as some of them contain special synaptic bodies (SBs) forming the so-called ribbon synapses. These SB-containing synapses are highly variable in number, exhibiting interspecies and intraspecies differences. As quantitative data on the incidence of SBs in the different rows of OHCs are lacking and as some of the above differences may have a circadian basis, in the present study SBs were counted in guinea pigs killed at different times of the day and night. In the second turn of the cochlea, synapses with perpendicular and parallel SBs were distinguished. Perpendicular SBs, but not parallel SBs, were significantly higher in number in the third row. Circadian changes were observed for perpendicular SBs, which were higher in number in the evening than in the morning. Circadian changes were absent in the parallel SBs of the OHCs and in the SBs of the inner hair cells. These results show that the afferent synapses of OHCs are rather complex, structurally as well as temporally; the functional significance of this finding remains to be elucidated.

Indirect evidence for the absence of electrotonic coupling between hair and support cells in the mammalian cochlea

Gap-junctional complexes link goldfish and lizard hair cells with adjacent supporting cells [Hama, J. Neurocytol. 9, 845–860 (1980); Nadol, Mulroy, Goodenough, and Weiss, Am. J. Anat. 147, 281–302 (1976)]. Lizard hair cells and support cells appear to be electrically coupled [Baden-Kristensen and Weiss, Hear. Res. 8, 295–315 (1982)]. In mammals, anatomical evidence suggests the absence of gap-junctional complexes between hair cells and support-cell neighbors, but suggestive membrane specializations have been observed [Gulley and Keese, J. Neurocytol. 5, 479–507 (1976); Nadol, Ann. Otol. 87, 70–80 (1978)]. Horseradish peroxidase marking and intracellular recording techniques were utilized to determine the origin (s) of sound-induced potential changes in supporting cells of the organ of Corti. Recordings were made in the low-frequency region of the adult guinea-pig cochlea, an in vivo preparation. Response characteristics of support cells to tone bursts at various sound levels and frequencies were compared with hair-cell responses (IHC and OHC) and potentials recorded in organ fluid spaces. Comparisons of the harmonic content of the responses, magnitude, and phase of the fundamental component, and the magnitude of de-response components suggest a lack of electrotonic coupling between mammalian hair cells and support cells. [Work supported by NIH Grant NS-08635.]

Actin filaments, stereocilia, and hair cells of the bird cochlea. V. How the staircase pattern of stereociliary lengths is generated.

The stereocilia on each hair cell are arranged into rows of ascending height, resulting in what we refer to as a "staircase-like" profile. At the proximal end of the cochlea the length of the tallest row of stereocilia in the staircase is 1.5 micron, with the shortest row only 0.3 micron. As one proceeds towards the distal end of the cochlea the length of the stereocilia progressively increases so that at the extreme distal end the length of the tallest row of the staircase is 5.5 micron and the shortest row is 2 micron. During development hair cells form their staircases in four phases of growth separated from each other by developmental time. First, stereocilia sprout from the apical surfaces of the hair cells (8-10-d embryos). Second (10-12-d embryos), what will be the longest row of the staircase begins to elongate. As the embryo gets older successive rows of stereocilia initiate elongation. Thus the staircase is set up by the sequential initiation of elongation of stereociliary rows located at increased distances from the row that began elongation. Third (12-17-d embryos), all the stereocilia in the newly formed staircase elongate until those located on the first step of the staircase have reached the prescribed length. In the final phase (17-d embryos to hatchlings) there is a progressive cessation of elongation beginning with the shortest step and followed by taller and taller rows with the tallest step stopping last. Thus, to obtain a pattern of stereocilia in rows of increasing height what transpires are progressive go signals followed by a period when all the stereocilia grow and ending with progressive stop signals. We discuss how such a sequence could be controlled.

Chapter 1 Ionic mechanisms in hair cells of the mammalian cochlea

Publisher Summary This chapter discusses the ionic mechanisms in hair cells of the mammalian cochlea. The sensory element common to all vertebrate inner ear structures is the hair cell. The ionic properties of inner and outer hair cells presented in the chapter are seen to be quite different. The role of the outer hair cell appears to be that of a device for boosting the motion of the basilar membrane near its best frequency. Outer hair cells maintained in extracellular medium are found to have a relatively low initial resting potential, typically –30 to –15 mV. The comparable measurements for inner hair cells are not known. The most surprising feature of isolated outer hair cells is that they are motile. The quasi-linear I-V curves for inner hair cells measured in vivo are inconsistent with the measurements on inner hair cells in vitro . The main function of such an arrangement is to reduce the metabolic load on such terminally differentiated cells.

Computer-aided three-dimensional reconstruction and morphometry of the outer hair cells of the guinea pig cochlea.

The outer hair cells and their nerve endings in the basal and third turns of the guinea pig cochlea were reconstructed three-dimensionally from serial thin sections by means of computer graphics, and morphometric data were obtained. The number of nerve endings in the third turn was two to three times greater than that in the basal turn. Many afferent and efferent terminals in the third turn did not demonstrate synaptic specialization. Presynaptic dense bodies were missing in the majority of outer hair cells in both basal and third turns. The morphologic arrangement of the subsurface cisternae and efferent fiber synapses on the side and base of the outer hair cells suggests a close functional relationship. The nerve fibers and cisternae may be involved in the contractile process of the cells. The volume of the outer hair cells in the first row of the basal turn was about 656 micron 3, and third turn, 1358 micron 3. The total count of mitochondria in the outer hair cells of the first row in the basal turn was 1425, and 1963 in the third turn. The density of mitochondria in the sensory cell in the basal turn was higher. The highest density was seen in the infranuclear region. The mitochondrial distribution patterns suggest that metabolic activity of the outer hair cells is higher in the basal turn than in the third turn and the energy requirement is greatest in the region close to nerve endings.

The morphology and physiology of hair cells in organotypic cultures of the mouse cochlea

Organotypic explant cultures were prepared from the cochleas of 1 to 3 day post-natal mice and maintained in vitro for up to 5 days. The hair cells retain morphological integrity for the duration of the culture period although they exhibit embryological features such as a kinocilium and additional microvilli on their apical surfaces. The resting membrane potentials of mouse inner hair cells (IHCs) in vitro are similar to those of guinea-pig IHCs in vivo but the membrane potentials of outer hair cells (OHCs) in the mouse cochlea in vitro are less polarized than the resting membrane potentials of OHCs in the basal turn of the guinea-pig cochlea in vivo. The voltage responses of IHCs and OHCs to sinusoidal displacements of their stereocilia are similar to each other in waveform and dynamic range, although the responses of IHCs are larger than those of OHCs. The relationship between transducer conductance and stereocilia displacement in IHCs and OHCs is non-linear and largely accounts for the depolarizing asymmetry of the voltage response. The receptor potentials of IHCs and OHCs reverse close to 0 mV indicating that the transducer conductance is non-selective for cations. The voltage responses of IHCs and OHCs to intracellular current injection rectify when the membrane potentials are more depolarized than about − 30 mV. This rectification is most pronounced in OHCs. OHCs also exhibit a time-dependent, voltage-sensitive conductance although they do not behave as electrical resonators.

The application of a capacitive probe technique for direct observation of electromechanical processes in the guinea pig cochlea.

The demonstration of evoked mechanical responses of the outer hair cells in the mammalian cochlea by indirect measurements introduces a new range of problems into direct mechanical measurements. Direct and indirect measurements indicate that the frequency spectra of evoked electromechanical responses may extend well into the range of audio frequencies, revealing a need to develop terminology and protocols for distinguishing evoked mechanical responses from the traditional traveling wave when both are apparently superimposed on the motion of the basilar membrane in the normally functioning cochlea. Details are presented of a frequency-modulation capacitive probe technique for measurement of vibrating structures of the guinea pig ear. Considerations include the design of the transducer, calibration, sensitivity, linearity, and sources of noise, as well as the influence of the technique upon the animal preparation, and in particular the issues associated with draining scala tympani for the measurement. Relative advantages and disadvantages of the technique are compared with salient features of other techniques currently available. In view of the apparent complexity of cochlear mechanics some preliminary experiments are required to elucidate some of the key questions about reverse-transduction processes in general. A "simple" first experiment is to test existence of any rectifying or motile response.

Computer-aided Three-dimensional Reconstruction of the Outer Hair Cells of the Guinea Pig Cochlea

Computer-aided Three-dimensional Reconstruction of the Outer Hair Cells of the Guinea Pig Cochlea

Modified staining technique for surface preparation of the organ of Corti

A simple technique for the staining of the inner ear tissues using toluidine blue and Ehrlich haematoxylin is described. The method is very convenient for histological evaluation of hair cell counts, for estimation of the number of scars in the reticular lamina, for observing the arrangement of stereociliary pattern and for the detection of traumatic changes after noise exposure. The method described proves to be very useful for histological examination of human temporal bones as well and makes possible an immediate evaluation of the hair cells of the human cochlea and direct correlation of the inner ear pathology with audiometric data.

Antagonistic effects of perilymphatic calcium and magnesium on the activity of single cochlear afferent neurons.

The dependence of the spontaneous and sound driven activity of single cochlear nerve fibers on the calcium and magnesium content of the perilymph was studied by perfusion of the perilymphatic space. It was possible to study these effects under steady-state conditions by continuously perfusing scala tympani at low rates while simultaneously recording from units in the chinchilla auditory nerve. Preparations were stable for many hours. As previously reported [Robertson and Johnstone (1979) Pflügers Arch. 380, 7-12], perfusion with solutions containing elevated concentrations of magnesium reduces both the spontaneous and driven activity. When calcium was eliminated from the perfusate, activity was completely abolished for stimuli with sound pressure levels below 100 dB. During partial blocks, a relatively frequency-independent threshold elevation was seen for frequencies well below the characteristic frequency (CF) of the unit, with greater elevations closer to CF. When the threshold elevation at CF was 30-40 dB, the width of the 'tip' portion of the tuning curve was reduced, resembling that of naturally-occurring units with low spontaneous rates of discharge. These effects are similar to that of raising the criterion for response during threshold measurement and are probably related to a frequency-dependent nonlinearity exhibited by the motion of the basilar membrane. The dynamic range for the growth of average rate with level was increased and saturation was shifted to higher stimulus levels during elevated magnesium perfusion. Raising the calcium content of the perfusate increased both spontaneous and driven rates, even in the saturated portion of the rate-intensity plot. Under these conditions, the response of the unit may more directly correspond to the intracellular potential of the presynaptic hair cell. It is argued that the primary site of divalent cation interaction is in the control of transmitter release. Inner hair cells of the mammalian cochlea apparently do not release transmitter in the absence of a calcium influx. The size of the pool of 'readily-available' transmitter appears to be influenced by divalent cations. Even though this synapse is probably specialized for the transmission of auditory signals, the mechanism of synaptic transmission is probably not fundamentally different from that of other well-characterized synapses.

The low-frequency response of inner hair cells in the guinea pig cochlea: Implications for fluid coupling and resonance of the stereocilia

AC receptor potentials within the inner hair cells of the basal turn of the guinea pig cochlea have been recorded for stimuli in the frequency range 20 Hz to 3200 Hz. Comparison of these potentials with potentials recorded in scala media suggests that the stereocilia of many inner hair cells are stimulated by the transverse velocity of the cochlear partition for very low frequency, but above a transition frequency in the range 400 Hz to 1000 Hz they become entrained with partition displacement. It is suggested that such a transition is probably a simple consequence of the fluid coupling that drives these cells, and that mechanical resonance of the free-standing stereocilia of the inner hair cells does not occur in the basal turn of the guinea pig. These results do not, however, preclude the possibility of mechanical resonance involving the stereocilia of the outer hair cells. The results also indicate that the bodies of these cells low-pass filter the intracellular receptor potential, with a cutoff frequency of approximately 1000 Hz.

The effect of chronic application of kanamycin on stereocilia and their tip links in hair cells of the guinea pig cochlea.

Albino guinea pigs were treated with kanamycin (400 mg/kg i.p.) daily for 10 days. After a 2-week recovery period their cochleae were analyzed by scanning electron microscopy. Attention was paid to those outer hair cells which had been less severely damaged. Stereocilia of the outer hair cells were often constricted at the root, and in some cases had become detached at the root. In stereocilia showing any degree of abnormality, the tip links were usually missing. However, in a few exceptional cases, tip links could remain on stereocilia showing other abnormalities, such as constriction at the root. Where hair cells were otherwise apparently unaffected, a much higher proportion of tip links remained, even on cells situated in an area of extensive hair cell loss. The results give further information on the process of kanamycin poisoning. They also suggest that substantial loss of tip links, and therefore perhaps of transduction, is one of the preliminary consequences of kanamycin poisoning.

Intercellular cross-linkages between the stereociliary bundles of adjacent hair cells in the guinea pig cochlea.

Hair cells of the guinea pig organ of Corti have been examined using high resolution scanning electron microscopy. In addition to the extensive array of cross-links between the stereocilia of individual hair cells which have been reported previously, we have seen examples of attachments between the stereocilia of both adjacent inner and adjacent outer hair cells. The implications of these observations are discussed.

Actin filaments, stereocilia, and hair cells of the bird cochlea: IV. How the actin filaments become organized in developing stereocilia and in the cuticular plate

In 8-day-old embryos stereocilia can be identified on the hair cells of the chick cochlea; within each is a small population of actin filaments which extend from the tip of the stereocilium to the apical cytoplasm of the cell. These filaments are not ordered in a regular way, however, and tend to be found near the lateral margins of the stereocilia with large spaces between adjacent filaments. By 9 days the spaces between adjacent filaments are reduced and there are regions where the crossover points of adjacent actin helices are in register even though in cross section the actin filaments do not lie on a regular lattice. By 10–11 days the actin filaments become progressively more crossbridged together and we can recognize in longitudinal section horizontal stripes caused by the periodicity of the crossbridges. In transverse section the filaments begin to lie on a hexagonal lattice. Each stereocilium, however, contains less than 100 actin filaments. Evidence is presented that once crossbridging is maximal and the filaments hexagonally packed (Days 11–12), the stereocilia increase in width by the orderly addition of actin filaments to the lateral margins of the existing filament bundle so that by Day 16 we find up to 400 filaments all packed on a hexagonal lattice. Thus there are two stages in bundle formation. In the first a small number of filaments condense into a hexagonally packed, crosslinked bundle. In the second, the bundle increases in diameter by addition of filaments to the periphery of the bundle in a process akin to crystal growth. From observations on the elongation of filaments in the rootlets and stereocilia, we conclude that rootlets grow by addition of subunits at the nonpreferred end while stereocilia elongate by addition to the preferred end. What makes this interesting is that these two modes of addition occur at different developmental times.

Ionic basis of membrane potential in outer hair cells of guinea pig cochlea

Mammalian hearing involves features not found in other species, for example, the separation of sound frequencies depends on an active control of the cochlear mechanics. The force-generating component in the cochlea is likely to be the outer hair cell (OHC), one of the two types of sensory cell through which current is gated by mechano-electrical transducer channels sited on the apical surface. Outer hair cells isolated in vitro have been shown to be motile and capable of generating forces at acoustic frequencies. The OHC membrane is not, however, electrically tuned, as found in lower vertebrates. Here we describe how the OHC resting potential is determined by a Ca2+-activated K+ conductance at the base of the cell. Two channel types with unitary sizes of 240 and 45 pS underlie this Ca2+-activated K+ conductance and we suggest that their activity is determined by a Ca2+ influx through the apical transducer channel, as demonstrated in other hair cells. This coupled system simultaneously explains the large OHC resting potentials observed in vivo and indicates how the current gated by the transducer may be maximized to generate the forces required in cochlear micromechanics.

Actin filaments, stereocilia, and hair cells of the bird cochlea: III. The development and differentiation of hair cells and stereocilia

The cochleae of chick embryos of 8 days of incubation until hatching (21 days) were examined by scanning electron microscopy. Unlike what one would expect from the literature, the total number of hair cells per cochlea (10,405 ± 529) is already determined and visible in a 10-day embryo and the growth of the cochlea is a result of the growth in size and surface area of the hair cells. We also find that the hair cells differentiate simultaneously throughout the cochlea and have followed the differentiation of individual hair cells throughout development. During development we find that the total number, hexagonal packing, and orientation of the stereocilia in each hair cell is determined early and accurately (9- to 10-day embryos). The stereocilia then begin to elongate in all the cells of the cochlea at approximately 0.5 μm/day. By Day 12 the tallest stereocilia in each cell are 1.5–1.8 μm long, the mature length for cells at the proximal end of the cochlea. At this point all stereocilia cease elongating, but those along the inferior edge gradually increase in width from 0.11 μm to maximally 0.19 μm in 17-day embryos. When the stereocilia on the inferior edge reach their mature width, widening ceases and the elongation of stereocilia in the distal hair cells begins again. When these stereocilia have attained their mature lengths, they stop growing. Thus elongation and widening of stereocilia are separated in time. During this period, 11 to 13 days, the shape of the tufts at the proximal end of the cochlea changes. This occurs because stereocilia in the front of each tuft are absorbed while others at the sides appear de novo. This rearrangement converts a circular bundle of stereocilia to a rectangular bundle.

Patterns of afferent synaptic contacts in the alligator lizard's cochlea.

Afferent synapses on both free-standing and tectorial hair cells in the alligator lizard's cochlea are described quantitatively. Semiserial sections were photographed with a transmission electron microscope. Hair cells together with their afferent nerves were reconstructed and morphometrically analyzed with the aid of a computer. Each afferent nerve forms many synapses with its hair cell. Tectorial afferents make more than twice the number of synapses with their hair cells as do free-standing afferents. This suggests a possible neuroanatomical basis for the physiological difference in synchrony reported in these two types of auditory nerve fibers; namely, the greater the number of synapses the better a fiber is able to follow faithfully the response of its hair cell.

GABA-like immunoreactivity in the squirrel monkey organ of corti

The distribution of γ-aminobutyric acid (GABA)-like immunoreactivity in the squirrel monkey organ of Corti was determined using an antiserum against GABA conjugated to bovine serum albumin. Immunoreactive labeling was seen in the region of the inner spiral bundle, the synaptic region below inner hair cells, in terminals contacting the basal part of outer hair cells, and in tunnel spiral fibers. Examples of each of these immunoreactive components could be observed in all cochlear turns. In the region of inner hair cells, immunoreactive labeling took the form of numerous small puncta randomly distributed below the base of the cells. In the region of outer hair cells, large globular immunoreactive structures reminiscent of terminal endings at the subnuclear level were observed. Since similar structures were seen at the base of outer hair cells in other cochleas processed for AChE, we conclude that GABA-like immunoreactivity was contained in efferent terminals which synapse on outer hair cells. These results strengthen previous evidence for the presence of GABA in the olivocochlear system of the mammalian cochlea.

The responses of inner and outer hair cells in the basal turn of the guinea-pig cochlea and in the mouse cochlea grown in vitro.

Until recently the responses of the mechanosensitive hair cells of the cochlea have been inferred from their morphology, morphological relationships with other structures in the cochlea, and by indirect electrophysiological measurements. With the advent of techniques for making intracellular recordings from hair cells in the cochleas of anaesthetised mammals it has been possible to measure the responses of hair cells to acoustic stimulation and to assess their roles in sensory transduction in the cochlea. Intracellular recordings of the responses of inner and outer hair cells in the basal turn of the guinea-pig cochlea show that they differ considerably from each other. The receptor potentials of inner hair cells are larger, predominantly depolarizing to low frequency tones and at their best frequencies (16-20 kHz) they generate depolarizing dc receptor potentials. Outer hair cells generate predominantly hyperpolarizing potentials to low frequency tones. They do not produce significant voltage responses at high frequencies except at high intensities when they generate slowly rising depolarizing potentials which are associated with loss of cochlear sensitivity. At low frequencies the receptor potentials of the inner hair cells phase lead those of the outer hair cell. Measurements of their frequency selectivity show that inner and outer hair cells are both sharply tuned. It is proposed that the responses of inner and outer hair cells are consistent with sensory and motor roles respectively in mechanoelectric transduction and that the outer hair cells are the site of an active mechanical process responsible for the frequency selectivity and sensitivity of the cochlea. Intracellular recordings from hair cells in the mouse cochlea maintained in vivo have provided a direct measure of the mechanosensitivity of cochlear hair cells (approximately 30 mV per degree of displacement of their stereociliary bundles) and indirect evidence that the transfer characteristics of the outer hair cells in vivo may be due to their mechanoelectrical interaction with the tectorial membrane. This is because the transfer characteristics of the inner and outer hair cells are similar in vitro in the absence of a tectorial membrane. Considerable importance is attributed to the shape of the transfer characteristics of the inner and outer hair cells. Changes in these characteristics during anoxia and following exposure to intense tones are associated with depolarization of the outer hair cells and loss of cochlear sensitivity and frequency selectivity. Current-voltage studies of hair cells in vivo show the inner and outer hair cells to be electrically nonlinear.(ABSTRACT TRUNCATED AT 400 WORDS)