Noise Stress Induces an Epidermal Growth Factor Receptor/Xeroderma Pigmentosum-A Response in the Auditory Nerve.

In response to toxic stressors, cancer cells defend themselves by mobilizing one or more epidermal growth factor receptor (EGFR) cascades that employ xeroderma pigmentosum-A (XPA) to repair damaged genes. Recent experiments discovered that neurons within the auditory nerve exhibit basal levels of EGFR+XPA co-expression. This finding implied that auditory neurons in particular or neurons in general have the capacity to mobilize an EGFR+XPA defense. Therefore, the current study tested the hypothesis that noise stress would alter the expression pattern of EGFR/XPA within the auditory nerve. Design-based stereology was used to quantify the proportion of neurons that expressed EGFR, XPA, and EGFR+XPA with and without noise stress. The results revealed an intricate neuronal response that is suggestive of alterations to both co-expression and individual expression of EGFR and XPA. In both the apical and middle cochlear coils, the noise stress depleted EGFR+XPA expression. Furthermore, there was a reduction in the proportion of neurons that expressed XPA-alone in the middle coils. However, the noise stress caused a significant increase in the proportion of neurons that expressed EGFR-alone in the middle coils. The basal cochlear coils failed to mobilize a significant response to the noise stress. These results suggest that EGFR and XPA might be part of the molecular defense repertoire of the auditory nerve.

The effect of progressive hearing loss on the morphology of endbulbs of Held and bushy cells.

Studies of congenital and early-onset deafness have demonstrated that an absence of peripheral sound-evoked activity in the auditory nerve causes pathological changes in central auditory structures. The aim of this study was to establish whether progressive acquired hearing loss could lead to similar brain changes that would degrade the precision of signal transmission. We used complementary physiologic hearing tests and microscopic techniques to study the combined effect of both magnitude and duration of hearing loss on one of the first auditory synapses in the brain, the endbulb of Held (EB), along with its bushy cell (BC) target in the anteroventral cochlear nucleus. We compared two hearing mouse strains (CBA/Ca and heterozygous shaker-2+/-) against a model of early-onset progressive hearing loss (DBA/2) and a model of congenital deafness (homozygous shaker-2-/-), examining each strain at 1, 3, and 6 months of age. Furthermore, we employed a frequency model of the mouse cochlear nucleus to constrain our analyses to regions most likely to exhibit graded changes in hearing function with time. No significant differences in the gross morphology of EB or BC structure were observed in 1-month-old animals, indicating uninterrupted development. However, in animals with hearing loss, both EBs and BCs exhibited a graded reduction in size that paralleled the hearing loss, with the most severe pathology seen in deaf 6-month-old shaker-2-/- mice. Ultrastructural pathologies associated with hearing loss were less dramatic: minor changes were observed in terminal size but mitochondrial fraction and postsynaptic densities remained relatively stable. These results indicate that acquired progressive hearing loss can have consequences on auditory brain structure, with prolonged loss leading to greater pathologies. Our findings suggest a role for early intervention with assistive devices in order to mitigate long-term pathology and loss of function.

Low Iron Diet Increases Susceptibility to Noise-Induced Hearing Loss in Young Rats.

We evaluated the role of iron deficiency (ID) without anemia on hearing function and cochlear pathophysiology of young rats before and after noise exposure. We used rats at developmental stages as an animal model to induce ID without anemia by dietary iron restriction. We have established this dietary restriction model in the rat that should enable us to study the effects of iron deficiency in the absence of severe anemia on hearing and ribbon synapses. Hearing function was measured on Postnatal Day (PND) 21 after induction of ID using auditory brainstem response (ABR). Then, the young rats were exposed to loud noise on PND 21. After noise exposure, hearing function was again measured. We observed the morphology of ribbon synapses, hair cells and spiral ganglion cells (SGCs), and assessed the expression of myosin VIIa, vesicular glutamate transporter 3 and prestin in the cochlea. ID without anemia did not elevate ABR threshold shifts, but reduced ABR wave I peak amplitude of young rats. At 70, 80, and 90 dB SPL, amplitudes of wave I (3.11 ± 0.96 µV, 3.52 ± 1.31 µV, and 4.37 ± 1.08 µV, respectively) in pups from the ID group were decreased compared to the control (5.92 ± 1.67 µV, 6.53 ± 1.70 µV, and 6.90 ± 1.76 µV, respectively) (p < 0.05). Moreover, ID without anemia did not impair the morphology hair cells and SGCs, but decreased the number of ribbon synapses. Before noise exposure, the mean number of ribbon synapses per inner hair cell (IHC) was significantly lower in the ID group (8.44 ± 1.21) compared to that seen in the control (13.08 ± 1.36) (p < 0.05). In addition, the numbers of ribbon synapses per IHC of young rats in the control (ID group) were 6.61 ± 1.59, 3.07 ± 0.83, 5.85 ± 1.63 and 12.25 ± 1.97 (3.75 ± 1.45, 2.03 ± 1.08, 3.81 ± 1.70 and 4.01 ± 1.65) at 1, 4, 7 and 14 days after noise exposure, respectively. Moreover, ABR thresholds at 4 and 8 kHz in young rats from the ID group were significantly elevated at 7 and 14 days after noise exposure compared to control (p < 0.05). The average number of young rat SGCs from the ID group were significantly decreased in the basal turn of the cochlea compared to the control (p < 0.05). Therefore, ID without anemia delayed the recovery from noise-induced hearing loss and ribbon synapses damage, increased SGCs loss, and upregulated prestin after noise exposure. Thus, the cochleae in rat pups with ID without anemia were potentially susceptible to loud noise exposure, and this deficit may be attributed to the reduction of ribbon synapses and SGCs.

The great auricular nerve in fetuses / El nervio auricular mayor en fetos

The great auricular nerve (GAN) is the largest branch of the superficial cervical plexus that winds around the posterior border of the sternocleidomastoid muscle, accompanied by the external jugular vein. Forty fetuses (right side: 40/80; left: 40/80) with gestational ages between 15 to 28 weeks were microdissected to document the anatomy of the GAN. The results obtained were classified as: i) Incidence and morphometry: GAN was present in 100 percent of the fetal specimens with average length on the right and left sides recorded as 12.65 +/- 2.14 mm and 12.55 +/- 2.82 mm respectively. ii) Course: GAN was located parallel to the transverse cervical nerve and the external jugular vein. Duplicate external jugular veins were observed in 5 percent (4/80) with GAN located anterior to one of the tributaries; iii) Branching Pattern: 16 percent (13/80) of specimens depicted a single branch. Duplicate branches in 67 percent (54/80) (referred to as Type I: anterior and posterior branches and 33 percent (26/80) referred to as Type II: anterior and posterior branches; iv) Variation: the transverse cervical nerve formed a communication with GAN, inferior to the parotid gland in 1 percent. The anatomical knowledge of the course, bifurcation pattern and variations of GAN may prevent complications during surgical procedures such as parotidectomies.

El nervio auricular mayor (NAM) es el ramo más grande del plexo cervical superficial que gira alrededor del margen posterior del músculo esternocleidomastoideo, acompañado de la vena yugular externa. Cuarenta fetos (lado derecho: 40/80; izquierdo: 40/80), con edades gestacionales de 15 a 28 semanas fueron microdisecados para describir la anatomía del NAM. Los resultados obtenidos se clasificaron en: i) Incidencia y morfometría: NAM estaba presente en el 100 por ciento de las muestras fetales con una longitud media de los lados derecho e izquierdo de 12,65+/-2,14mm y 12,55+/-2,82mm, respectivamente. ii) Curso: NAM se encuentra paralelo al nervio cervical transverso y la vena yugular externa. Duplicación de las venas yugulares externas se observaron en el 5 por ciento (4/80) con el NAM situado por delante de uno de los afluentes, iii) Patrón de ramificación: 16 por ciento (13/80) de las muestras presentaba una solo ramo. Ramos duplicados en el 67 por ciento (54/80) de Tipo I, ramos anterior y posterior y, el 33 por ciento (26/80) Tipo II, ramos anterior y posterior, y iv) Variación: el nervio cervical transverso formando una comunicación con NAM, inferior a la glándula parótida en el 1 por ciento. El conocimiento anatómico del curso, patrón de bifurcación y variaciones del NAM pueden prevenir las complicaciones durante los procedimientos quirúrgicos como la parotidectomía.

Expression of myelin basic protein in the human auditory nerve - an immunohistochemical and comparative study.

OBJECTIVE: The aim of this study is to analyse the expression and distribution of myelin basic protein (MBP or Myelin A1 protein) in the human spiral ganglion and auditory nerve. MATERIALS AND METHODS: Cryostat sections were made from freshly fixed human cochlear specimens removed at surgery in patients with life-threatening petro-clival meningiomas compressing the brain stem. The sections were subjected to immunohistochemistry using antibodies against MBP, S-100 and Tubulin. The immunoreaction was documented using laser confocal microscopy. RESULTS: Type I spiral ganglion nerve somata (SGN) were surrounded by so-called "satellite glial cells" (SGCs) that lacked expression of MBP consistent with earlier light and electron microscopic findings indicating that these cells are non-myelinating. S-100 labeling showed that the SGCs form a continuous network in the apical region. CONCLUSIONS: The pattern of myelination in human spiral ganglion is different from that in other species' spiral ganglion. The striking differences in myelin outline should be investigated further in combination with its influence on signal coding and preservation properties in man.

The border between the central and the peripheral nervous system in the cat cochlear nerve: a light and scanning electron microscopical study.

The transition between the central (CNS) and peripheral nervous system (PNS) in cranial and spinal nerve roots, referred to here as the CNS-PNS border, is of relevance to nerve root disorders and factors that affect peripheral-central regeneration. Here, this border is described in the cat cochlear nerve using light microscopical sections, and scanning electron microscopy of the CNS-PNS interfaces exposed by fracture of the nerve either prior to or following critical point drying. The CNS-PNS border represents an abrupt change in type of myelin, supporting elements, and vascularization. Because central myelin is formed by oligodendrocytes and peripheral myelin by Schwann cells, the myelinated fibers are as a rule equipped with a node of Ranvier at the border passage. The border is shallower and smoother in cat cochlear nerve than expected from other nerves, and the borderline nodes are largely in register. The loose endoneurial connective tissue of the PNS compartment is closed at the border by a compact glial membrane, the mantle zone, of the CNS compartment. The mantle zone is penetrated by the nerve fibers, but is otherwise composed of astrocytes and their interwoven processes like the external limiting membrane of the brain surface with which it is continuous. The distal surface of the mantle zone is covered by a fenestrated basal lamina. Only occasional vessels traverse the border. From an anatomical point of view, the border might be expected to be a weak point along the cochlear nerve and thus vulnerable to trauma. In mature animals, the CNS-PNS border presents a barrier to regrowth of regenerating nerve fibers and to invasion of the CNS by Schwann cells. An understanding of this region in the cochlear nerve is therefore relevant to head injuries that lead to hearing loss, to surgery on acoustic Schwannomas, and to the possibility of cochlear nerve regeneration.

Cell transplantation to the auditory nerve and cochlear duct.

We have developed a technique to deliver cells to the inner ear without injuring the membranes that seal the endolymphatic and perilymphatic chambers. The integrity of these membranes is essential for normal hearing, and the technique should significantly reduce surgical trauma during cell transplantation. Embryonic stem cells transplanted at the internal auditory meatal portion of an atrophic auditory nerve migrated extensively along it. Four-five weeks after transplantation, the cells were found not only throughout the auditory nerve, but also in Rosenthal's canal and the scala media, the most distal portion of the auditory nervous system where the hair cells reside. Migration of the transplanted cells was more extensive following damage to the auditory nerve. In the undamaged nerve, migration was more limited, but the cells showed more signs of neuronal differentiation. This highlights an important balance between tissue damage and the potential for repair.

Experimental vibratory damage of the inner ear.

The aim of the experiment was to determine the effect of whole-body vibration on the inner ear. The investigations were carried out on 40 guinea pigs, subjected to sinusoidal vibration (10 Hz/5 mm/1.4 g rms) for 1 to 6 months in a noiseless apparatus. Cochlear microphonic measurements were done with a phase-sensitive detection technique for the levels 70, 80 and 90 dB and the frequencies of 0.26, 0.5, 1 and 2 kHz from the apex of the cochlea and for 4 and 8 kHz from the region of the round window. Analysis of 1,440 measurements suggested the possibility of damage appearing in the upper turnings of the cochlea. The subsequent morphological analysis was based on the estimation of the state of the hair cells (a three-degree scale of injury) in a Zeiss DSM 950 scanning microscope and of the structure of the fibers of the acoustic nerve in a Zeiss EM 900 transmission microscope. Vibration-induced changes were seen in all the examined inner ears of the experimental groups. Hair-cell damage was more often seen in the region of the apex, spreading gradually in the direction of the base and from the circumference (outer hair cells of the third row) to the modiolus. The most characteristic vibrational changes of the acoustic nerve fibers occurred in 100% of the examined myelin sheaths and were visible as decreases in their electrodensity. The changes in both the assessed elements of the inner ear appeared simultaneously but independently and were directly connected with the duration of the experiment. The results obtained allow an explanation of the mechanism of hearing loss in persons subjected to whole-body vibration. The damages done to the inner ear structures may cause a worsening of hearing there, especially in the low and medium frequencies.

A model of the electrically excited human cochlear neuron. I. Contribution of neural substructures to the generation and propagation of spikes.

Differences in neural geometry and the fact that the soma of the human cochlear neuron typically is not myelinated are reasons for disagreements between single fiber recordings in animals and the neural code evoked in cochlear implant patients. We introduce a compartment model of the human cochlear neuron to study the excitation and propagation process of action potentials. The model can be used to predict (i) the points of spike generation, (ii) the time difference between stimulation and the arrival of a spike at the proximal end of the central axon, (iii) the vanishing of peripherally evoked spikes at the soma region under specific conditions, (iv) the influence of electrode positions on spiking behavior, and (v) consequences of the loss of the peripheral axon. Every subunit of the cochlear neuron is separately modeled. Ion channel dynamics are described by a modified Hodgkin--Huxley model. Influence of membrane noise is taken into account. Additionally, the generalized activating function is introduced as a tool to give an envision of the origin of spikes in the peripheral and in the central axon without any knowledge of the gating processes in the active membranes. Comparing the reactions of a human and cat cochlear neuron, we find differences in spiking behavior, e.g. peripherally and centrally evoked spikes arrive with a time difference of about 400 mus in man and 200 mus in cat.

Morphological relationships of peptidergic and noradrenergic nerve terminals to olivocochlear neurones in the rat.

In the rat, the outer hair cells in the cochlea receive direct synaptic input from neurones in the ventral nucleus of the trapezoid body. These so-called medial olivocochlear neurones exert an inhibitory influence on the cochlear neural output. Electrophysiological in vitro studies suggest that the activity of medial olivocochlear neurones may be affected by a variety of neuropeptides as well as noradrenaline, but anatomical confirmation of direct synaptic input is still lacking. We have investigated, at the light microscopical level, the morphological relationships between terminals containing noradrenaline, substance P, cholecystokinin and leu-enkephalin, and medial olivocochlear neurones in the rat. A retrograde tracer was injected into the cochlea to label medial olivocochlear neurones and a double labelling immunocytochemical method was used to visualise the retrograde tracer as well as the neurotransmitters within each brain section. Light microscopical analysis revealed nerve endings containing substance P, cholecystokinin and leu-enkephalin in close apposition to the dendrites of medial olivocochlear neurones, and nerve endings containing dopamine-beta-hydroxylase, a marker for noradrenaline, in close contact with the somata as well as dendrites of medial olivocochlear neurones. Although the technique cannot prove the existence of functional synaptic contacts, the results are broadly consistent with electrophysiological data and suggest a direct input to medial olivocochlear neurones from substance P, cholecystokinin, leu-enkephalin and noradrenaline-containing neural pathways. Differences in the densities and spatial distribution of the various neuropharmacological inputs suggest differences in the relative strengths and possible roles of these diverse inputs to the olivocochlear system.

The limitations of hearing preservation in acoustic neuroma surgery: histological study of the interface between the eighth cranial nerve and the tumor.

The interface between the eighth cranial nerve and acoustic neuroma was investigated by light microscopy and immunohistochemistry in paraffin sections in 13 patients, and in Epon sections in 19 patients. The cochlear nerve was severely invaded by tumor cells in two of six specimens examined, moderately invaded in two specimens, and not invaded in two specimens. Gliosis was frequently found within the cochlear and vestibular nerve. Hemangioma-like tissue was occasionally found attached to the cochlear and vestibular nerve adjacent to the tumor. These findings were considered to be related to difficulty in hearing preservation in some patients although acoustic neuroma surgery was successful.

Involvement of cochlear nerve in acoustic tumours.

With conventional light and transmission electronmicroscopy we studied 10 cases of acoustic nerve tumour, 3 of which proved to be instances of von Recklinghausen neurofibroma and 7 of schwannoma. Schwannomas were not found to infiltrate the cochlear nerve. Hearing loss, if present in cases of schwannoma, could be related to non-specific lesions of the uninfiltrated cochlear nerve in the vicinity of the vestibular nerve tumour. Only neurofibromas were found to infiltrate the cochlear nerve. Distinction between tumour infiltration and non-specific lesion could be made by electron microscopy.

Glycine receptor immunoreactivity in the ventral cochlear nucleus of the guinea pig.

Glycine appears to be a major inhibitory neurotransmitter in the cochlear nucleus. In order to determine more precisely the distribution of glycinergic synapses, we have studied the immunocytochemical distribution of the glycine postsynaptic receptor. Two monoclonal antibodies were used, Gly Rec Ab 2, which recognizes the 48kD polypeptide and Gly Rec Ab 7, which primarily recognizes the 93kD subunit of the glycine receptor complex. At the light microscopic level, glycine receptor immunoreactivity was found throughout the ventral cochlear nucleus with a punctuate distribution often found outlining large cell bodies. Indistinguishable patterns of staining were obtained with the two antibodies. Ultrastructural localization was done with Gly Rec Ab 7 because immunoreactivity remained after fixation with glutaraldehyde containing solutions. At the ultrastructural level, immunoreactivity was concentrated at postsynaptic sites on dendrites and cell bodies. In the anteroventral cochlear nucleus, neurons identified as spherical cells contained numerous inmunoreactive synapses on their cell bodies, whereas most immunoreactive synapses on stellate cells were on their proximal dendrites. In the posteroventral cochlear nucleus, neurons identified as octopus cells were immunoreactive on their cell bodies and proximal dendrites. In the granule cell layer, immunoreactivity was found only in the neuropile. Throughout the ventral cochlear nucleus, glycine receptor immunoreactivity was found postsynaptic to terminals containing flattened synaptic vesicles as well as those containing oval/pleomorphic synaptic vesicles.

The polypeptide PEP-19 is a marker for Purkinje neurons in cerebellar cortex and cartwheel neurons in the dorsal cochlear nucleus.

This light and electron microscopic immunocytochemical study shows that the polypeptide PEP-19, a presumptive calcium binding protein specific to the nervous system, represents an excellent marker for cerebellar Purkinje cells and dorsal cochlear nucleus (DCoN) cartwheel cells. The polypeptide clearly reveals the entire populations of both types of neurons, including their complete dendritic and axonal arborizations. Other PEP-19 containing neurons in the two regions display weak immunoreactivity restricted to the cell body or to cell body and principal dendrites. Electron microscopic localization of PEP-19-like immunoreactivity reveals similarities between this polypeptide, parvalbumin, and a 28K vitamin D-dependent calcium binding protein. However, calmodulin, which is expressed in both Purkinje and granule cells, may differ from PEP-19. Similarities between the organization of the cerebellar cortex and the DCoN superficial layers have been known for some time, with several types of neurons in one system having their presumed homologue in the other. These data provide further support for the proposed structural and functional homology between Purkinje and cartwheel neurons, and establishes PEP-19 as a useful marker for examining degeneration of these two neuronal populations in murine cerebellar mutants.

Functional and histological findings in acoustic neuroma.

27 patients with acoustic neuroma were operated on by translabyrinthine tumor removal. At operation 8th nerve compartments were identified and removed for study. Results of the audiological test battery and vestibular tests were compared with histological state of the nerve specimens studied. The tumor appears only seldom to invade the cochlear nerve in cases with 'cochlear' hearing loss, but in the majority of cases with 'retrocochlear' hearing loss the cochlear nerve is invaded.

Ultrastructural features of spiral ganglion cells. A study of patients with hearing loss of varying origins.

Fresh specimens of the acoustic nerve that were obtained during vestibular neurectomy or acoustic neuroma removal were studied for structure of the spiral ganglion cells. These were large, round or oval cells with many cytoplasmic organelles and were surrounded by Schwann cells. Schwann cells formed a single layer or perikaryal sheath and the cells were thus unmyelinated. In one case the specimen contained part of the spiral ganglion, but in six others spiral ganglion cells were found outside of Rosenthal's canal, among the myelinated nerve fibers. In three cases the ear was deaf due to cochlear insult, but the nerve fibers and ganglion cells showed no retrograde degeneration. The neurophysiologic features of the cochlear nerve should be considered because unmyelinated ganglion cells may have a different conduction capacity in comparison with thickly myelinated cells.

[Oncorna-like virus particles in the cochlear spiral ganglion of guinea pigs (author's transl)]. / Oncorna-ähnliche Viruspartikel im Ganglion spirale cochleae des Meerschweinchens.

INTRODUCTION: Virus particles in spiral ganglion cells of the guinea pig have been grouped with herpes viruses. The purpose of this study was to evaluate this classification of the viruses and their influence on cell morphology. MATERIAL AND METHODS: 20 healthy, adult guinea pigs from different breeds were studied. The spiral ganglion was serially sectioned and examined electron micoscopically. RESULTS AND DISCUSSION: All examined spiral ganglions of several guinea pig populations from different breeds showed intracytoplasmic viruses in some granular spiral ganglion cells. According to their localization and morphology we classify these viruses with the oncorna virus group. This is not in agreement with the classification of other authors. Apparently there is a world-wide latent viral infection in guinea pigs. The accumulation of lysosomal-like vacuoles in vicinity to the viruses indicates an increased local lysosomal activity of the infected ganglion cells. Considering the otherwise normal ultrastructure of the infected cells an additional influence of these viruses on the intracellular metabolism can neither be demonstrated nor denied.