Expression patterns of estrogen receptors in the central auditory system change in prepubertal and aged mice.

Estrogens are important in the development, maintenance and physiology of the CNS. Several studies have shown their effects on the processing of hearing in both males and females, and these effects, in part, are thought to result from regulation of the transcription of genes via their classical estrogen receptor (ER) pathway. In order to understand the spatiotemporal changes that occur with age, we have studied the expression of ERs in the central auditory pathway in prepubertal and aged CBA mice with immunohistochemistry. In prepubertal mice a clear dichotomy was noted between the expression of ERα and ERß. ERß-positive neurons were found in the metencephalon whereas the majority of ERα was found in mesencephalon, diencephalon or the telencephalon. In the aged animals a different pattern of ER expression was found in terms of location and overall intensity. These age-induced changes in the expression pattern were generally not uniform, suggesting that region-specific mechanisms regulate the ERs' age-related expression. Neither the prepubertal nor the aged animals showed sex differences in any auditory structure. Our results demonstrate different age-dependent spatial and temporal changes in the pattern of expression of ERα and ERß, suggesting that each ER type may be involved in distinct roles across the central auditory pathway in different periods of maturation.

The remarkable cochlear amplifier.

This composite article is intended to give the experts in the field of cochlear mechanics an opportunity to voice their personal opinion on the one mechanism they believe dominates cochlear amplification in mammals. A collection of these ideas are presented here for the auditory community and others interested in the cochlear amplifier. Each expert has given their own personal view on the topic and at the end of their commentary they have suggested several experiments that would be required for the decisive mechanism underlying the cochlear amplifier. These experiments are presently lacking but if successfully performed would have an enormous impact on our understanding of the cochlear amplifier.

Prevalence and characteristics of hearing problems in a working and non-working Swedish population.

BACKGROUND: Hearing problems are among the top 10 most common burdens of disease and are projected to be become even more common by the year 2030. The aim of the present study was to give a current assessment of the prevalence of communication difficulties because of hearing loss and tinnitus, in the general Swedish working and non-working populations in relation to sex, age, socioeconomic status (SES) and noise exposure. How prevalence is affected by SES has not been previously established. METHODS: A total of 18 734 individuals were invited to participate in the study, of which 11 441 (61%) enrolled. Of the participants, 9756 answered the questionnaire for those who work and 1685 answered the version for non-workers. FINDINGS: The most important findings are that 31% in the working population and 36% in the non-working population report either hearing loss or tinnitus or both. The prevalence of hearing problems increases with age, is higher among men and persons with low self-rated SES, and covaries with exposure to noise at work. Severe hearing problems are already present in men and women under 40 years of age who are exposed to work-related noise. INTERPRETATION: Prevalence of hearing problems is far more common than previously estimated and is associated with SES and noise exposure history. Hearing problems have a gradual onset that can take years to become recognised. In order to proactively intervene and prevent this deleterious, yet avoidable handicap, statistics need to be regularly updated.

Estrogen receptors in the central auditory system of male and female mice.

The estrogen receptors in the central auditory system of male and female mice were characterized using immunocytochemical methods. Estrogen receptors alpha and beta (ERalpha, ERbeta) were localized predominantly in the ventral cochlear nucleus, nucleus of the trapezoid body, the lateral- and medio-ventral periolivary nuclei, the dorsal lateral lemniscus, and the inferior colliculus. The medial geniculate nucleus was negative for both ERalpha and ERbeta whereas the auditory cortex was positive for ERalpha. The lateral superior olive, the ventral lateral lemniscus and the central nucleus of the inferior colliculus expressed only ERbeta. The differential localization of ERalpha and ERbeta may indicate distinct roles for these two receptors in auditory processing. No major differences in the pattern, number or intensity of receptor expression was found between male and female animals. The comprehensive anatomic map that is constructed for ERalpha and ERbeta in the central auditory pathway will be a useful foundation to elucidate the complexity of estrogen actions in the auditory system.

Differential activation of mitogen-activated protein kinases and brain-derived neurotrophic factor after temporary or permanent damage to a sensory system.

Functional and morphological differences between temporary (TTS) and permanent (PTS) hearing loss induced by acoustic trauma are well characterized whereas molecular differences remain to be elucidated. A comparative analysis of the expression of the phosphorylated forms of extracellular signal-regulated kinase (ERK1/2), c-jun-N-terminal kinases 1/2 (JNK1/2) and p38 in the mouse cochlea after acoustic trauma resulting in either a temporary or permanent damage is presented. In the acute phase of PTS an upregulation of phosphorylated p38, JNK1/2, and ERK1/2 was found while in the acute phase of TTS a downregulation of phospho-p38 occurred and no immediate change of pJNK1/2 and pERK1/2 was noted. After a 24 h recovery from TTS JNK1/2 and ERK1/2 was activated while the expression of phospho-p38 was downregulated. In contrast PTS group showed complete recovery to control values for all three MAPKs by 24 h post. The level of brain-derived neurotrophic factor (BDNF), a potent otoprotective agent, was elevated after both types of acoustic trauma but the elevation after permanent trauma was of a longer duration. The expression of BDNF receptor's TrkB (truncated form) was downregulated only after permanent hearing loss. Thus, temporary and permanent hearing loss demonstrate different expression patterns and temporal aspects of MAPK, BDNF and TrkB in the cochlea. The results of this study will help reveal the cellular mechanisms underlying hearing loss induced by acoustic trauma.

The signal transduction pathway for the dopamine D1 receptor in the guinea-pig cochlea.

Dopamine released from lateral efferent fibers modulates the activity of the auditory nerve, but the signaling mechanism by which this is mediated is not known. The present study investigated the signal transduction pathway for the dopamine D1 receptor in the guinea-pig cochlea. D1 receptor immunolabeling was localized to the spiral ganglia neurons and at the base of the inner hair cells. Western immunoblotting on whole cochlear preparations revealed positive bands for the D1 receptor and for dopamine and the cyclic AMP-regulated phosphoprotein. The amplitude of the compound action potential was enhanced in the presence of the D1 receptor agonist, SKF 38393, an effect that was abolished by H89, a protein kinase A inhibitor. Conversely, SKF 83566, a D1 receptor antagonist decreased the amplitude of compound action potential, while forskolin, a protein kinase A activator prevented this effect. Furthermore, it was found that the level of glutamate receptor 1 phosphorylation at the protein kinase A site (Ser845) was increased by the D1 agonist, but decreased by D1 antagonist. Our results provide evidence that the D1 receptor is localized in the spiral ganglion neurons as well as the nerve endings under the inner hair cells and they can modulate auditory nerve function. One signal transduction pathway of D1 receptor in the auditory nerve is via protein kinase A-mediated glutamate receptor 1 phosphorylation.

The distribution and the modulation of tyrosine hydroxylase immunoreactivity in the lateral olivocochlear system of the guinea-pig.

It was previously shown that tyrosine hydroxylase (TH) immunoreactivity in the terminals of the lateral efferents of the cochlea is decreased by acoustic trauma and that sound preconditioning counteracted this decrease [Hear Res 174 (2002) 124]. Here we identify those neurons in the lateral olivocochlear system (LOC) in the brainstem that regulates the peripheral expression of TH in the cochlea. By employing retrograde tracing techniques, dextran-labeled neurons were found predominantly in the ipsilateral LOC system including lateral superior olive (LSO), and the surrounding periolivary regions (dorsal periolivary nucleus [DPO], dorsolateral periolivary nucleus [DLPO], lateral nucleus of trapezoid body [LNTB]). Employing immunocytochemistry, it was found that a control group had 35% of the ipsilateral LOC neurons positively stained with TH. Of the total population of TH neurons, 77% were double-stained (TH and dextran) in the LOC system. Acoustic trauma decreased the number of TH positive neurons in the LSO and the surrounding DLPO, and caused a reduction of TH fiber immunolabeling in these regions. Changes were not found in the DPO or the LNTB after acoustic trauma. Sound conditioning protected against the decrease of TH immunolabeling by acoustic trauma and increased the fiber staining for TH in the LSO and DLPO, but not in the DPO or the LNTB. These results provide evidence that TH positive neurons are present in the LOC system in the guinea-pig. It is now demonstrated that protection against acoustic trauma by sound conditioning has a central component that is governed by TH in the LSO and the surrounding periolivary DLPO region.

The total number of neurons and calcium binding protein positive neurons during aging in the cochlear nucleus of CBA/CaJ mice: a quantitative study.

The quantitative stereological method, the optical fractionator, was used for determining the total number of neurons and the total number of neurons immunostained with parvalbumin, calbindin-D28k (calbindin), and calretinin in the dorsal and posteroventral cochlear nucleus (DCN and PVCN) in CBA/CaJ (CBA) mice during aging (1-39 months old). CBA mice have only a modest sensorineural pathology late in life. An age-related decrease of the total number of neurons was demonstrated in the DCN (r=-0.54, P<0.03), while the total number of neurons in the PVCN did not show any significant age-related differences (r=0.16, P=0.57). In the DCN 5.5% of neurons were parvalbumin positive in the very old (30-39 months) mice, vs. 2.2% in the 1 month old mice. In the DCN 3% of the neurons were calbindin immunopositive in the 30-39 months mice compared to 1.9% in the 1 month old group. In the PVCN, 20% of the neurons in the very old mice were parvalbumin immunopositive, compared to 12% in the young mice. Calbindin did not show any significant age-related differences in the PVCN. The total number of calretinin immunopositive neurons both in the DCN and PVCN did not show any significant change with increasing age. In conclusion, the total neuronal number in the DCN and PVCN was age-related and region-specific. While the neuronal number in the DCN and PVCN was decreased or unchanged, respectively, the calcium binding protein positive neuronal number showed a graded increase during aging in a region-specific and protein-specific manner.

Peripheral cell loss related to calcium binding protein immunocytochemistry in the dorsal cochlear nucleus in CBA/CaJ mice during aging.

The influence of cochlear hair cell and spiral ganglia neuron loss on calcium binding protein immunoreactivity (calretinin, parvalbumin and calbindin) in the dorsal and posteroventral cochlear nuclei (DCN and PVCN) in CBA/CaJ (CBA) mice during aging (1-39 months) was determined. Since calcium binding proteins have buffering properties against calcium overload, they may have a protective role during aging. It is shown that the percentage of calretinin- and parvalbumin-immunopositive neurons in the DCN showed a statistically significant positive correlation with inner hair cell loss, outer hair cell loss, and spiral ganglion cell loss. A correlation was also found between aging and the auditory periphery, and calcium binding proteins in the DCN. These findings imply that the pathophysiological state of the auditory periphery may influence the neuronal homeostasis in the dorsal cochlear nucleus.

Short-term adaptation in the peripheral auditory system is related to the AMPA receptor.

The role of glutamate receptors was investigated by infusing N-methyl-D-aspartate (NMDA) or alpha-amino-3-hydroxy-5-methyl-isoxazol-propionate (AMPA) into the guinea pig cochlea. Auditory brainstem response thresholds and forward masking were used to determine auditory sensitivity. In the presence of 330 microM NMDA, the auditory brainstem response (ABR) thresholds were elevated by 20-30 dB at 2 kHz and 8 kHz, and the slopes of the forward masking curves were not significantly different from controls. When a high concentration of NMDA (15 mM) was used, ABR thresholds were elevated by 40-50 dB at 2 kHz and 8 kHz and the slopes of the forward masking curves were significantly decreased. In contrast, when AMPA (150 microM) was infused, ABR thresholds were elevated by 20-35 dB at 2 and 8 kHz and the slopes of the forward masking curves were significantly decreased from the control group. When the concentration of AMPA was decreased (100 microM). ABR thresholds were not significantly altered but the slopes of the forward masking curves were significantly decreased from control values. The present study suggests that AMPA receptors play a significantly more important role in short-term adaptation than NMDA receptors.

Succinic dehydrogenase histochemistry as an early marker for hair cell pathology.

Density measurements of succinic dehydrogenase (SDH) activity were obtained from the inner and outer hair cells on surface preparations obtained from the guinea pig cochlea. Guinea pigs were exposed to noise (3.85 kHz, 120 dB SPL, 22.5 min) and sacrificed 0, 4 or 24 h after the exposure. By 4 h after exposure, the first- and second-row outer hair cells already demonstrated an altered SDH activity. By 24 h after exposure, a significant decrease in SDH staining in both the inner and outer hair cells at a distance of 10-12 mm from the cochlear apex was demonstrated. After a 1-month recovery period, scanning electron microscopy confirmed the main lesion site to be at a distance of 10-12 mm. In addition, Hensen's cells (supporting cells) at a distance of 10-12 mm from the apex were intensely stained by SDH after noise exposure, indicating an increase in oxidative metabolism. SDH staining in the Hensen's cells from the unexposed cochleae was not found. In conclusion, our findings suggest that the early use of SDH histochemistry can predict later permanent damage to the organ of Corti.

Noise-induced aspartate and glutamate efflux in the guinea pig cochlea and hearing loss.

Aspartate and glutamate were monitored in the scala tympani of the guinea pig cochlea using in vivo microdialysis before and during noise exposure. Moderate level broad band noise [105 dB sound pressure level (SPL), 30 min] neither altered the levels of aspartate or glutamate, nor auditory brainstem response (ABR) thresholds. High level noise exposure (135 dB SPL, 30 min) caused a large increase in aspartate (330%), a smaller increase in glutamate (150%), and a permanent ABR threshold shift of 60-75 dB between 2.0 and 12.5 kHz. Morphological analysis of the cochlea revealed a collapse of supporting structures, swelling of the afferent dendrites under the inner hair cells, and outer hair cell loss. Pretreatment with the NMDA antagonist, MK 801 (1 mg/kg body weight, i.p.) 1 h before noise exposure protected the afferent dendrites from swelling but did not protect the collapse of supporting structures, outer hair cell loss, or auditory thresholds. In conclusion, the noise-induced increase in aspartate and glutamate release in the cochlea and the protective effect of NMDA antagonism suggest that these two neurotransmitters are involved in noise-induced hearing loss.

Complementary roles of neurotrophin 3 and a N-methyl-D-aspartate antagonist in the protection of noise and aminoglycoside-induced ototoxicity.

Recent progress has been made regarding the prevention of hearing loss. However, the complete protection of both hair cells and spiral ganglion neurons, with restored function, has not yet been achieved. It has been shown that spiral ganglion neuronal loss can be prevented by neurotrophin 3 (NT3) and hair cell damage by N-methyl-D-aspartate (NMDA) receptor antagonists. Here we demonstrate that the combined treatment with MK801, a NMDA antagonist, and NT3 protect both cochlear morphology and physiology from injury. Pretreatment with MK801 prevented hearing loss and the dendrites of the spiral ganglion neurons from swelling after noise-induced damage. The acute phase of insult with the aminoglycoside antibiotic amikacin resulted in swollen afferent dendrites beneath the inner hair cells. The chronic phase resulted in complete hair cell loss and near-complete loss of spiral ganglion neurons. This damage caused a near-complete loss of hearing sensitivity as displayed by elevated (>90-dB sound pressure levels) auditory brainstem response thresholds. The treatment of amikacin-exposed animals with MK801 gave only a partial protection of hearing. However, the combined treatment with NT3 and MK801 in the amikacin-comprised ear resulted in improved mean hearing within 20 dB of normal. Furthermore, hair cell loss was prevented in these animals and spiral ganglion neurons were completely protected. These results suggest that the NMDA antagonist MK801 protects against noise-induced excitotoxicity in the cochlea whereas the combined treatment of NT3 and MK801 has a potent effect on preserving both auditory physiology and morphology against aminoglycoside toxicity.

Characterization of hearing in an X,0 'Turner mouse'.

Turner's syndrome is due to total (45,X) or partial (mosaicism) loss of one X-chromosome. The main features are short stature, ovarian dysgenesis with no estrogen production and infertility. In addition to ear and hearing disorders, middle ear problems including acute/serous otitis media and chronic middle ear disease are frequent. Sensorineural hearing loss is often seen with a dip in the mid-frequencies and also an early high frequency loss. In this study, middle-and inner-ear pathology was characterized using physiological and morphological techniques in a 'Turner mouse' that has been generated with the chromosomal aberration X,0. Otitis media was found in some of these X,0 animals, a symptom that is seldom found in control animals. The auditory brainstem responses (ABR) of the Turner mouse showed a progressive hearing loss in the high frequency region that exceeded the normal age-related hearing loss of control mice and increased latencies of the first ABR wave. Outer hair cell loss was apparent in the cochlear basal turn of Turner mice. Decreases in the amplitude of distortion product otoacoustic emissions were correlated with the loss of ABR threshold sensitivity. These results indicate that hearing problems in the Turner mouse seems to be of cochlear origin with an eighth nerve component. This Turner mouse model appears to have ear and hearing problems quite similar to humans and can therefore be used as a model to determine the auditory pathology underlying this syndrome.

Sound stimulation increases calcium-binding protein immunoreactivity in the inferior colliculus in mice.

The numerical density of calbindin D-28k and parvalbumin immunopositive neurons in the inferior colliculus (IC) in mice was increased after sound stimulation. An increased number of calbindin positive neurons was found in the deep layers of the external cortex (EC) and particularly in the dorsal cortex (DC) and commissural nucleus (NCO). An increase of parvalbumin positive neurons was found in the EC, central nucleus (ICC) and DC, but not in the NCO. The increased immunoreactivity related to sound exposure suggests the appearance of neurons which express these proteins after sound stimulation. The up-regulation of calcium-binding proteins in these neurons may be due to their protective role against overstimulation, their response to a higher auditory metabolic activity, or increasing effect of excitatory inputs after noise-induced hearing loss.

Medial olivocochlear efferent terminals are protected by sound conditioning.

Synaptophysin immunoreactivity was used as a marker for the olivocochlear efferent system that innervates the outer hair cells of the cochlea. An intense noise exposure at either 6.3 kHz or 1.0 kHz caused a significant reduction in anti-synaptophysin immunoreactivity within the 8-6 mm or 14-11 mm distance from the round window, respectively. In the region of the main lesion, the reduction in synaptophysin immunoreactivity for both the 6.3 and 1.0 kHz exposures correlated well with outer hair cell loss. In regions peripheral to the main lesion, some remnants of efferent nerve endings could remain even when their associated outer hair cells were missing. Pre-treatment with a low level sound conditioner (either at 6.3 tone or 1.0 kHz) effectively reduced the efferent and outer hair cell pathology induced by the 6.3 and 1.0 kHz intense noise exposures, respectively. The results demonstrate the feasibility of using anti-synaptophysin immunoreactivity as an effective means of quantifying pathological alterations to the medial cochlear efferent terminals throughout the cochlea. Furthermore, the results show that sound conditioning significantly reduces damage to the efferent terminals.

Neurotrophins, NMDA receptors, and nitric oxide in development and protection of the auditory system.

Neurotrophic factors are secreted peptides that when interacting with specific classes of membrane receptors activate intracellular signaling cascades that prevents neuronal death during embryonic development. The establishment of gene-targeted null mutant mice for the neurotrophic factors, brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT3) has led to the discovery that they are crucial trophic factors for the survival of auditory and vestibular neurons during development. BDNF is the major survival factor for vestibular ganglion neurons, while NT-3 only support a small number of these neurons. In the cochlea, auditory type I neurons require NT3 for their survival, whereas type II neurons depend on BDNF. With this information at hand recent progress has been made regarding the prevention of aminoglycoside-induced hearing loss in the adult guinea pig. These results and the mechanisms leading to hair cell damage are discussed in this paper.

Modulating calbindin and parvalbumin immunoreactivity in the cochlear nucleus by moderate noise exposure in mice. . A quantitative study on the dorsal and posteroventral cochlear nucleus.

The number of calbindin D-28k and parvalbumin immunoreactive (IR) neurons were characterized on sections from the cochlear nucleus, dorsal cochlear nucleus (DCN) and posteroventral cochlear nucleus (PVCN) using two-dimensional quantification. After noise exposure (6-12 kHz, 2 h, at either 80 dB SPL or 103 dB SPL), the number of calbindin and parvalbumin immunoreactive neurons increased in CBA/CBA mice. Quantitative analysis of calbindin-IR in the PVCN did not show a statistically significant difference between any of the groups, whereas statistically significant differences in calbindin-IR were found in the DCN for the 103 dB and 80 dB group compared to the control group, and 103 dB compared to the 80 dB group, respectively. A statistically significant increase in the number of parvalbumin-IR neurons in the PVCN and the DCN was evident in the 103 dB and 80 dB group compared to the control group, and in the 103 dB compared to the 80 dB group. The data indicate that increasing sound stimulation causes a graded increase in the expression of calcium-binding protein immunoreactivity in the DCN and PVCN neurons and neuropil. This increase of protein expression is due to increased positive immunoreactivity in 'silent' neurons. These findings implicate that these neurons have the possibility to react against trauma and display calbindin or parvalbumin as a rescue event. The ability to map sound-induced calcium-binding protein changes in auditory neurons may be useful in future studies designed for detecting early patterns of neurodegeneration and neuroprotection in the central auditory pathway.

Receptor potential characteristics during direct stereocilia stimulation of isolated outer hair cells from the guinea-pig.

The receptor potential as a function of stimulus amplitude and frequency was studied with the patch-clamp technique in isolated outer hair cells (OHCs) with a length ranging from 30 to 87 microm during direct mechanical stimulation of the stereocilia. The amplitude and frequency of the stimulation were varied from 125 nm to 2 microm and from 100 Hz to 2.5 kHz, respectively. The mean resting membrane potential before stimulation was -64.25 +/- 1.4 mV (mean +/- SE, n = 26). Irrespective of the frequency used, stereocilia stimulation produced a combination of AC and DC responses, and both components showed saturation with increasing stimulation. Frequency responses appeared to be a function of intensity and resembled a low-pass filter with a time constant ranging from 0.2 to 2.0 ms. With increasing stereocilia stimulation, the relative contribution of high frequencies to the AC component decreased, suggesting a decrease of the corner frequency. The saturated amplitude of the AC component for low-frequency stimulation (100 Hz) was proportional to cell length and increased with a mean rate of 0.014 mV microm(-1). A relationship between the DC response of the receptor potential and the pre-stimulus membrane potential was found. Recordings with more negative membrane potentials had greater DC components, while more depolarized recordings demonstrated smaller DC components. These fluctuations seemed to be defined by the interaction between the probe and stereocilia bundle and could be in the range of the transfer function for each cell.