Jean W.T. Smolders

A physiological place-frequency map of the cochlea in the CBA/J mouse.

Genetically manipulated mice have gained a prominent role in in vivo research on development and function of the auditory system. A prerequisite for the interpretation of normal and abnormal structural and functional features of the inner ear is the exact knowledge of the cochlear place-frequency map. Using a stereotaxic approach to the projection site of the auditory nerve fibers in the cochlear nucleus, we succeeded in labelling physiologically characterized auditory nerve afferents and determined their peripheral innervation site in the cochlea. From the neuronal characteristic frequency (CF) and the innervation site in the organ of Corti a place-frequency map was established for characteristic frequencies between 7.2 and 61.8 kHz, corresponding to locations between 90% and 10% basilar membrane length (base = 0%, apex = 100%, mean length measured under the inner hair cells 5.13 mm). The relation between normalized distance from the base (d) and frequency (kHz) can be described by a simple logarithmic function: d(%) = 156.5-82.5 x log(f), with a slope of 1.25 mm/octave of frequency. The present map, recorded under physiological conditions, differs from earlier maps determined with different methods. The simple logarithmic place-frequency relation found in the mouse indicates that mice are acoustic generalists rather than specialists.

Basilar membrane motion in the pigeon measured with the Mössbauer technique.

Vibration measurements were made of the basilar membrane (BM), limbi and columella footplate (CFP) of pigeon using the Mössbauer technique. Recordings were located at 0.23-1.33 mm from the basal end of the BM. The existence of a travelling wave mode, propagating from base to apex, was established for papillae in apparently good physiological condition. For these papillae the characteristic frequency (CF) of the BM isovelocity (0.08 mm X s-1) response was an exponential function of distance with a frequency mapping constant of 0.91 +/- 0.10 mm (equivalent to 0.63 +/- 0.07 mm X oct-1); BM CF at the base was 5.95 +/- 0.65 kHz. Travelling wave motion was not demonstrated for papillae in poor physiological condition; tonotopy of BM CF was still evident, although the correlation with distance was less (1.08 +/- 0.30 mm X oct-1; 4.35 +/- 0.73 kHz at the base). BM motion was linear and the isovelocity responses were less sensitive and less sharp than single unit threshold tuning curves: for papillae in good physiological condition the SPL at BM CF at 0.08 mm X s-1 was 51 +/- 6 dB SPL; Q10 dB was 1.24 +/- 0.38; high- and low-frequency slopes were 20 +/- 6 dB X oct-1 and -14 +/- 4 dB X oct-1, respectively. The response of the BM relative to the CFP for papillae in good physiological condition was reminiscent of a second order resonant system with damping constant of 0.33 +/- 0.06 and group delay at BM CF of 0.89 +/- 0.36 periods.

Effects of temperature on the properties of primary auditory fibres of the spectacled caiman,Caiman crocodilus (L.)

SummaryThe effect of temperature on the response properties of primary auditory fibres in caiman was studied. The head temperature was varied over the range of 10–35 ° C while the body was kept at a standard temperature of 27 °C (Ts). The temperature effects observed on auditory afferents were fully reversible. Below 11 °C the neural firing ceased.The mean spontaneous firing rate increased nearly linearly with temperature. The slopes in different fibres ranged from 0.2–3.5 imp s−1 °C−1. A bimodal distribution of mean spontaneous firing rate was found (<20 imp s−1 and >20 imp s−1 at Ts) at all temperatures.The frequency-intensity response area of the primary fibres shifted uniformly with temperature. The characteristic frequency (CF) increased nearly linearly with temperature. The slopes in different fibres ranged from 3–90 Hz °C−1. Expressed in octaves the CF-change varied in each fibre from about O.14oct °C−1 at 15 °C to about 0.06 oct °C−1 at 30 °C, irrespective of the fibres CF at Ts. Thresholds were lowest near Ts. Below Ts the thresholds decreased on average by 2dB°C−1, above Ts the thresholds rose rapidly with temperature. The sharpness of tuning (Q10db) showed no major change in the temperature range tested.Comparison of these findings with those from other lower vertebrates and from mammals shows that only mammalian auditory afferents do not shift their CF with temperature, suggesting that a fundamental difference in mammalian and submammalian tuning mechanisms exists. This does not necessarily imply that there is a single unifying tuning mechanism for all mammals and another one for non-mammals.

Functional Recovery in the Avian Ear after Hair Cell Regeneration

Trauma to the inner ear in birds, due to acoustic overstimulation or ototoxic aminoglycosides, can lead to hair cell loss which is followed by regeneration of new hair cells. These processes are paralleled by hearing loss followed by significant functional recovery. After acoustic trauma, functional recovery is rapid and nearly complete. The early and major part of functional recovery after sound trauma occurs before regenerated hair cells become functional. Even very intense sound trauma causes loss of only a proportion of the hair cell popu- lation, mainly so-called short hair cells residing on the abneural mobile part of the avian basilar membrane. Uncoupling of the tectorial membrane from the hair cells during sound overexposure may serve as a protection mechanism. The rapid functional recovery after sound trauma appears not to be associated with regeneration of the lost hair cells, but with repair processes involving the surviving hair cells. Small residual functional deficits after recovery are most likely associated with the missing upper fibrous layer of the tectorial membrane which fails to regenerate after sound trauma. After aminoglycoside trauma, functional recovery is slower and parallels the structural regeneration more closely. Aminoglycosides cause damage to both types of hair cells, starting at the basal (high frequency) part of the basilar papilla. However, functional hearing loss and recovery also occur at lower frequencies, associated with areas of the papilla where hair cells survive. Functional recovery in these low frequency areas is complete, whereas functional recovery in high frequency areas with complete hair cell loss is incomplete, despite regeneration of the hair cells. Permanent residual functional deficits remain. This indicates that in low frequency regions functional recovery after aminoglycosides involves repair of nonlethal injury to hair cells and/or hair cell-neural synapses. In the high frequency regions functional recovery involves regenerated hair cells. The permanent functional deficits after the regeneration process in these areas are most likely associated with functional deficits in the regenerated hair cells or shortcomings in the synaptic reconnections of nerve fibers with the regenerated hair cells. In conclusion, the avian inner ear appears to be much more resistant to trauma than the mammalian ear and possesses a considerable capacity for functional recovery based on repair processes along with its capacity to regenerate hair cells. The functional recovery in areas with regenerated hair cells is considerable but incomplete.

A functional map of the pigeon basilar papilla: correlation of the properties of single auditory nerve fibres and their peripheral origin

The purpose of the investigation was to correlate the functional properties of primary auditory fibres with the location of appertaining receptor cells in the avian basilar papilla. The functional properties of 425 single afferent fibres from the auditory nerve of adult pigeons were measured. The peripheral innervation site of 39 fibres was identified by intracellular labelling and correlated with the fibres functional properties. Mean spontaneous firing rate (SR, 0.1-250/s) was distributed monomodally (mean: 91 +/- 47/s) but not normally. Characteristic frequencies (CFs) were in the range of 0.02-4 kHz. SR, threshold at CF (4-76 dB SPL) and sharpness of tuning (Q10 dB, 0.1-8.8) varied systematically with CF. For a given CF there was a strong correlation of threshold and Q10 dB and of threshold and SR. Labelled fibres innervated different hair cell types over 93% of the length and 97% of the width of the basilar papilla. The majority of fibres innervated hair cells located between 30 and 70% distance from the apex and 0 and 30% distance from the neural edge of the papilla. CFs are mapped tonotopically from high at the base to low at the apex of the papilla, with a mean mapping constant of 0.63 +/- 0.05 mm/octave (in vivo). The highest CF at the base extrapolates to 5.98 +/- 1.17 kHz. The lowest CF mapped at the apex is 0.021 kHz. From the data, together with data from mechanical measurements (Gummer et al., 1987), a frequency-place function of the pigeon papilla was calculated. Transverse gradients of threshold at CF and of Q10 dB were observed across the width of the papilla. Thresholds were lowest and sharpness of tuning was highest above the neural limbus at a distance of 23% from the neural edge of the papilla. Hair cells in this sensitive strip are the tallest and narrowest ones across the width of the papilla. They are packed most densely and receive the largest number of afferent fibres. Fibres innervating (mostly short) hair cells on the free basilar membrane were spontaneously active and responsive to sound. Their Q10 dB was less than average but their sensitivity and SR were comparable to the mean population values. It is concluded that functional properties change gradually not only along the length but also across the width of the pigeon basilar papilla. The results support the idea that sharp frequency tuning of avian primary auditory fibres involves tuning mechanisms supplementary to the tuning of the free part of the basilar membrane.

Hearing Loss in Athyroid Pax8 Knockout Mice and Effects of Thyroxine Substitution

Pax8–/– mice do not develop thyroid follicular structures and thus provide an ideal animal model to study the consequences of congenital hypothyroidism. Despite their athyroidism, Pax8–/– mice survive up to postnatal day 21 (P21). No auditory brain stem responses (ABR) to sound could be recorded in these animals at 130 dB SPL, even at P21, when hearing reaches adult sensitivity in control mice. Abnormalities in the outer and middle ear structures were found in a considerable percentage of Pax8–/– animals. Maturation of the inner ear appeared delayed by about 1 week with respect to euthyroid controls. Hearing of adult Pax8–/– mice could be nearly normalized by early postnatal substitution with thyroxine (T4), but structural and functional restoration of hearing was incomplete. Even when T4 substitution was initiated at P1, ABR thresholds, measured at 6 weeks of age or more, were increased by about 20 dB, and each day of delay in the start of T4 substitution resulted in an additional threshold loss of about 4 dB. The most prominent structural deficit in Pax8–/– animals in which T4 substitution was started at P8 or later was an abnormally thick tectorial membrane. In these late-substituted animals, disarray of stereovilli from inner and outer hair cells was observed and also outer hair cell loss was found, predominantly in the basal part of the cochlea. The degree of structural disorder increased the later T4 substitution was initiated. The structural and functional consequences of postnatal athyroidism observed in Pax8–/– mice are largely in agreement with and extend those data obtained from hypothyroid animal models in which hypothyroidism was induced by goitrogenic agents (methimazole, propylthiouracil) or animal models with disrupted genes for the TSH receptor or the thyroid hormone receptors. The hearing loss and also the recovery effect by T4 substitution in Pax8–/– mice is larger than that in the other models. Although Pax8–/– mice are born by euthyroid Pax8+/– dams, the Pax8–/– phenotype could not be completely restored by immediate postnatal T4 substitution, indicating that some deficits are the consequence of prenatal T4 deficiency of the offspring.

Synchronized responses of primary auditory fibre-populations in Caiman crocodilus (L.) to single tones and clicks

Measurements of the responses to tones and clicks were made from single primary auditory fibres of the caiman. The distribution of the amplitude and phase of the fundamental component of the response rate modulation over the best frequencies of the fibres is comparable to that reported in the cat, despite the fact that the basilar membrane in caiman is only 4.5 mm long. However, much higher intensities are needed in the caiman (75-85 dB SPL) than reported in the cat (20 dB SPL) to obtain systematic distributions of the phase of the responses, probably due to the larger scatter of the phase responses in the caiman. The slopes of the phase distributions are very similar to those in cat. Single unit phase responses as a function of stimulus frequency at 85 dB SPL can be approximated by one, or in fibres with low best frequency, two straight lines. At lower intensities the deviation of the phase-frequency responses from a straight line increases as the group delay at the best frequency becomes larger. The shortest latencies of click responses are obtained with rarefaction clicks. Group delay estimates obtained from the responses to clicks and from the straight line approximations of the phase-frequency responses are related in a way expected for linear filter systems and accurately predict the measured distributions of the phase of the responses over the neural best frequency. The obtained group delays and click latencies in the caiman are very similar to those reported by other workers in the cat, the squirrel monkey and the treefrog, despite large morphological and probably functional differences of their inner ears. The click latencies are also very similar to those in the pigeon. The results are consistent with the existence of a mechanical travelling wave reported previously on the basilar membrane of the caiman, but at the same stimulus level the phase characteristic of the present single unit responses is steeper and the wave length estimates from the neural population phase distributions are shorter than those observed directly in the motion of the basilar membrane. Since the neural responses are an indirect estimate of the basilar membrane motion it cannot be decided whether the difference between neural and mechanical data is due to deterioration of the basilar membrane responses during the direct measurements or whether the basilar membrane response is sharpened by additional tuning mechanisms.

Mechanics of a single-ossicle ear: I. The extra-stapedius of the pigeon.

The motion of the conical peak of the tympanic membrane (TM) at the tip of the extra-stapedius (ES) and of the columella footplate (CFP) were measured in the pigeon using the Mössbauer technique. The dimensions of middle-ear structures were measured in some of the experimental animals. The averaged velocity response at the ES for frequencies of 0.25-2.378 kHz was that of a second order, mass and stiffness controlled, resonant system with resonant frequency of 1.2 kHz and Q3 dB of 1.2. The mean velocity amplitude at resonance was 3.7 mms-1 at 100 dB SPL, which is approximately equal to the theoretical value of 3.5 mms-1 required for maximum energy transfer from a uniform plane acoustic wavefront in air. For the frequency regions 0.125-0.25 kHz and 2.378-5.657 kHz, the mean amplitude slopes for the velocity at the ES were 2 dB oct-1 and -3 dB oct-1, respectively. Above 5.657 kHz there was considerable inter-animal variation in the ES velocity responses. The direction of motion at the ES was frequency dependent above 1 kHz. For frequencies up to 1 kHz the ratio of CFP to ES velocity was independent of frequency; the mechanical lever ratio was 2.7, which was attributed to the geometry of the middle ear. At these frequencies the total transformer ratio for the middle ear, expressing the ratio of fluid pressure at the CFP to sound pressure at the ES, was estimated to be 35 dB.

Mechanics of the basilar membrane in Caiman crocodilus

Vibration measurements were made at a number of positions near the proximal (basal) end of the basilar membrane, and on the columella footplate, of Caiman crocodilus using a capacitive probe. The measurements established the existence of a mechanical travelling wave in this species. They showed no significant change of mechanical tuning with temperature, and were highly significantly different from previous reports of neural temperature sensitivity (Smolders, J. and Klinke, R. (1984): J. Comp. Physiol. 155, 19-30). Thus the neural sensitivity to temperature change appears not to depend upon basilar membrane mechanics. One interpretation of this is that the basilar membrane passively precedes an active temperature-sensitive filter. It was also found that the limbus supporting the basilar membrane had a measurable, but unturned, vibration and that the effect of draining scala tympani for the measurements was to increase the basilar membrane tuning frequency by a factor of about 1.5.

Preferred intervals in birds and mammals: A filter response to noise?

Quasi-periodic spontaneous activity (preferred intervals, PIs) has been reported from avian primary auditory afferents. In mammals, PIs have not been reported, as yet. As the length of PIs is close to 1/characteristic frequency, it has been suggested that this type of spontaneous activity indicates particular mechanisms in avian inner ear transduction. However, the present paper shows that pigeon auditory fibres possessing preferred intervals in their spontaneous activity always belong to the most sensitive and the most sharply-tuned fibres recorded. This leads to the assumption that preferred intervals are the response of narrow-band filters to noise. This view is supported by three additional findings: (i) Near-threshold noise provokes PIs in avian fibres that show no spontaneous PIs. (ii) Similarly, PIs can also be evoked in mammalian (gerbil) auditory afferents by low level noise. (iii) Phase-locking of auditory afferents can be achieved by sound stimuli 10-20 dB below rate threshold. It is argued that no conclusions may be drawn from the presence of PIs about the nature of the underlying filter.