Trauma-specific insults to the cochlear nucleus in the rat.

The effect of acoustic overstimulation on the neuronal number of the cochlear nucleus (CN) was investigated by using unbiased stereological methods in rats. We found that, after 9 weeks of recovery, neurons in the anteroventral cochlear nucleus (AVCN) degenerated, whereas those in the posteroventral and dorsal cochlear nuclei (PVCN and DCN) were preserved. The noise trauma induced near complete loss of the outer hair cells throughout the cochlea, and the inner hair cells were preserved only in the more apical regions. This pattern of selective loss of AVCN neurons in this study was different from trauma induced by auditory deafferentation by mechanical compression of auditory neurons. In contrast to noise trauma, mechanical compression caused loss of neurons in the PVCN and DCN. After 5 weeks of recovery from mechanical compression, there was no loss of inner or outer hair cells. These findings indicate that auditory deprivation, induced by different experimental manipulations, can have strikingly different consequences for the central auditory system. We hypothesized that AVCN neuronal death was induced by excitotoxic mechanisms via AMPA-type glutamate receptors and that excitatory neuronal circuits developed after acoustic overstimulation protected the PVCN and DCN against neuronal death. The results of the present study demonstrate that hearing loss from different etiologies will cause different patterns of neuronal degeneration in the CN. These findings are important for enhancing the performance of cochlear implants and auditory brainstem implants, because diverse types of hearing loss can selectively affect neuronal degeneration of the CN.

Silencing p27 reverses post-mitotic state of supporting cells in neonatal mouse cochleae.

The post-natal cochlear mammalian epithelium have no capacity to proliferate in tissue, however, dissociated supporting cells exhibit the ability to divide and trans-differentiate into new hair cells in vitro, with this process found to be correlated with the downregulation of the cyclin-dependent kinase inhibitor p27(kip1). Here we show that knockdown of p27(kip1) with short hairpin RNA-expressing vectors results in the cell-cycle reentry of post-mitotic supporting cells in the post-natal mouse cochleae ex vivo. The p27(kip1)-knockdown cells incorporated BrdU, and then divided into two daughter cells. However, there was also activation of the apoptotic pathway in some supporting cells. These results indicate that the use of RNA interference to target p27(kip1) is an effective strategy for inducing cell-cycle reentry in post-mitotic supporting cells in the post-natal mammalian cochleae, although additional manipulations of the supporting cells are required to achieve hair cell regeneration.

Development of the Reading Cognitive Test Kyoto (ReaCT Kyoto) for Early Detection of Cognitive Decline in Patients with Hearing Loss.

BACKGROUND: Early detection of cognitive decline allows timely intervention to delay progression of dementia. However, current cognitive evaluation tools often include items delivered via verbal forms of instruction, which can cause poor performance in patients with hearing loss. OBJECTIVE: To develop and validate a cognitive screening battery, the Reading Cognitive Test Kyoto (ReaCT Kyoto), comprising test items given through non-verbal instruction. METHODS: A cross-sectional and multi-center study was conducted in the three medical institutes. ReaCT Kyoto was designed to evaluate domains of "registration," "repetition," "delayed recall," "visuospatial recognition," "orientation in time and place," and "executive function." The Japanese version of the Mini-Mental State Examination Test (MMSE-J) and ReaCT Kyoto were applied by experienced psychotherapists. Concurrent validity was evaluated between the ReaCT Kyoto Test and MMSE-J and between the ReaCT Kyoto Test and physician-diagnosed dementia. RESULTS: ReaCT Kyoto was validated in a sample of 115 participants. The mean age of subjects was 81.0±6.4 years, and the sample comprised 53.0% females. The area under the receiver operating curves was 0.95 for detecting physician-diagnosed dementia. When classifying patients in accordance with presence or absence of hearing loss, the AUCs were 0.93 and 0.97 for those with and without hearing loss, respectively. With a cut-off score of < 29 points for suspected dementia, ReaCT Kyoto correctly classified 90.4% of the subjects as belonging to the group with or without physician-diagnosed dementia. CONCLUSION: ReaCT Kyoto provides an appropriate solution for detection of cognitive impairment in persons with or without hearing loss.

Isolation and Characterization of Mammalian Otic Progenitor Cells that Can Differentiate into Both Sensory Epithelial and Neuronal Cell Lineages.

The mammalian inner ear mediates hearing and balance and during development generates both cochleo-vestibular ganglion neurons and sensory epithelial receptor cells, that is, hair cells and support cells. Cell marking experiments have shown that both hair cells and support cells can originate from a common progenitor. Here, we demonstrate the lineage potential of individual otic epithelial cell clones using three cell lines established by a combination of limiting dilution and gene-marking techniques from an embryonic day 12 (E12) rat otocyst. Cell-type specific marker analyses of these clonal lines under proliferation and differentiation culture conditions demonstrate that during differentiation immature cell markers (Nanog and Nestin) were downregulated and hair cell (Myosin VIIa and Math1), support cell (p27Kip1 and cytokeratin) and neuronal cell (NF-H and NeuroD) markers were upregulated. Our results suggest that the otic epithelium of the E12 mammalian inner ear possess multipotent progenitor cells able to generate cell types of both sensory epithelial and neural cell lineages when cultured under a differentiation culture condition. Understanding the molecular mechanisms of proliferation and differentiation of multipotent otic progenitor cells may provide insights that could contribute to the development of a novel cell therapy with a potential to initiate or stimulate the sensorineural repair of damaged inner ear sensory receptors. Anat Rec, 303:451-460, 2020. © 2019 American Association for Anatomy.

"Replacement of diseased auditory neurons by cell transplantation".

The auditory nerve is an important target in hearing restoration research along with the hair cells. Although there are several potentially useful therapeutic options to rebuild lost hearing, cell transplantation is a very realistic option. Cells can be infused into the auditory nerve without compromising the auditory brainstem responses and damaging the membranous labyrinth. The final fate of transplanted cells may be determined by the intrinsic molecular program and the extracellular guidance cues. The first factor may be largely decided by the type of donor cell used and the second factor can be modified by the application of various molecules. Our recent experiments using ontogenetic-stage/region-restricted precursors and embryonic stem cells suggest that donor cells at later development stages seemed to have more mature intrinsic molecular programs to guide them more precisely and efficiently to the final expected destination. We discuss the critical interactions between the extracellular molecules such as myelin-derived inhibitory molecules expressed after CNS injury and the intracellular actin dynamics regulated by Rho GTPases in relation to the regeneration of the auditory neurons.

Potential of embryonic stem cell-derived neurons for synapse formation with auditory hair cells.

Recent studies have indicated that embryonic stem cells (ESCs) can be a source for the replacement of spiral ganglion neurons (SGNs), auditory primary neurons, and neurite projections from ESC-derived neurons to auditory sensory epithelia. However, the potential of ESC-derived neurons for synapse formation with auditory hair cells (HCs) has not been elucidated. The present study therefore aimed to examine the ability of ESC-derived neurons to form synaptic connections with HCs in vitro. Mouse ESC-derived neural progenitors expressing enhanced green fluorescence protein (EGFP) were cocultured with explants of cochlea sensory epithelia obtained from postnatal day 3 mice. After a 7-day culture, neurites of ESC-derived neurons predominantly elongated toward inner hair cells (IHCs), which play a crucial role in sound transmission to SGNs. Immunohistochemical analyses revealed the expression of synapsin 1 and synaptophysin in the nerve endings of ESC-derived neurons adjacent to IHCs, indicating the formation of synaptic connections. Transmission electron microscopy demonstrated synaptic contacts between nerve endings of ESC-derived neurons and IHCs. The present findings show that ESC-derived neurons can make synaptic connections with IHCs.

Association of cholesterol efflux capacity with plasmalogen levels of high-density lipoprotein: A cross-sectional study in chronic kidney disease patients.

BACKGROUND AND AIM: Current research suggests that dysfunctional high-density lipoprotein (HDL) with low cholesterol efflux capacity may accelerate atherosclerosis, particularly in chronic kidney disease (CKD). We previously reported that serum levels of plasmalogens closely correlated with HDL concentration, and could serve as a novel biomarker for atherosclerosis. In the present study, we analyzed the association of cholesterol efflux capacity of HDL with clinical and biochemical parameters, including plasmalogens, in CKD patients. METHODS: We enrolled 24 mild-to-moderate CKD patients (CKD-3-4) and 33 end-stage renal disease (ESRD) patients nearing hemodialysis (CKD-5), and assessed physiological atherosclerotic scores, cholesterol efflux capacity, and plasmalogens levels in HDL. Furthermore, the effect of plasmalogen on cholesterol efflux capacity of HDL was examined by in vitro studies with re-constituted HDL (rHDL) and HDL prepared from CKD-5 patient (ESRD-HDL) with additional phospholipids. RESULTS: There were significant differences in many parameters between the two groups. In particular, plasmalogens levels and cholesterol efflux capacity of HDL were significantly reduced in the CKD-5 group compared to those in the CKD-3-4 group (-35.1%, p < 0.001, -36.8%, p < 0.001, respectively). Multivariate linear regression analyses revealed that ethanolamine plasmalogen levels of HDL were independently associated with cholesterol efflux capacity (p = 0.045) and plaque scores (p = 0.035). In vitro studies also indicated that additional plasmalogens augmented cholesterol efflux ability of HDL. CONCLUSIONS: High plasmalogens concentrations in HDL may correlate with acceleration of cholesterol efflux and their decreased levels may promote atherosclerosis in advanced CKD patients.

Investigation of 5-HT3 receptor-triggered serotonin release from guinea-pig isolated colonic mucosa: a role of PYY-containing endocrine cell.

The effect of a 5-HT3 receptor-selective agonist SR57227A was investigated on the outflow of 5-hydroxytryptamine (5-HT) from isolated muscle layer-free mucosal preparations of guinea-pig colon. The mucosal preparations were incubated in vitro and the outflow of 5-HT from these preparations was determined by high-performance liquid chromatography with electrochemical detection. SR57227A (100µM) produced a tetrodotoxin-resistant and sustained increase in the outflow of 5-HT from the mucosal preparations. The SR57227A-evoked sustained 5-HT outflow was completely inhibited by the 5-HT3 receptor antagonist ramosetron (1µM). The neuropeptide Y1 receptor antagonist BIBO3304 (100nM) partially inhibited the SR57227A-evoked sustained 5-HT outflow, but the Y2 receptor antagonist BIIE0246 (1µM) or the glucagon-like peptide-1 (GLP-1) receptor antagonist exendin-(9-39) (1µM), showed a minimal effect on the SR57227A-evoked sustained 5-HT outflow. In the presence of BIBO3304 (100nM) and exendin-(9-39) (1µM), SR57227A (100µM) failed to produce a sustained increase in the outflow of 5-HT. The Y1 receptor agonist [Leu31, Pro34]-neuropeptide Y (10nM), but not GLP-1-(7-36) amide (100nM), produced a sustained increase in the outflow of 5-HT. We found that 5-HT3 receptor-triggered 5-HT release from guinea-pig colonic mucosa is mediated by the activation of 5-HT3 receptors located at endocrine cells (enterochromaffin cells and peptide YY (PYY)-containing endocrine cells). The activation of both Y1 and GLP-1 receptors appears to be required for the maintenance of 5-HT3 receptor-triggered 5-HT release. It is therefore considered that 5-HT3 receptors located at colonic mucosa play a crucial role in paracrine signaling between enterochromaffin cells and PYY-containing endocrine cells.

Recovery of hair cell function after damage induced by gentamicin in organ culture of rat vestibular maculae.

Here, we report the functional and morphological evidence of hair cell recovery after damages induced by gentamicin (GM) in cultured explants of rat vestibular maculae. We evaluated mechano-electrical transduction (MET) function in hair cells, by measuring Ca(2+) responses in the explants with fura-2 when hair bundles were stimulated. After the MET testing, hair bundles were observed in high resolution by scanning electron microscopy, or by fluorescence microscopy after staining with phalloidin-FITC (fluorescent isothiocyanate). In the control culture, the number of hair bundles on the explants gradually decreased, and the percentage of explants showing Ca(2+) responses decreased and disappeared after 17 days in culture. Following GM (1-2 mM) treatment, most of the hair bundles were eliminated initially, but the hair bundles gradually increased in number during culture. Short hair bundle-like structures emerged in the areas where hair bundles had been completely lost. Consistent with the morphological observations, Ca(2+) responses disappeared after GM treatment, and they gradually recovered to a peak 13-17 days after treatment and were even induced at 17 days or more in culture. Furthermore, cells accumulated FM1-43, a dye permeable through the MET channel, when Ca(2+) responses recovered after GM treatment. Application of steroid hormone increased the percentage of explants showing MET activity, and enhanced the recovery of MET after GM treatment. We investigated Ki-67 immunoreactivity to detect cell proliferation and TUNEL staining to detect apoptotic cell death. Ki-67 immunoreactivity was negative after GM treatment, however TUNEL staining was positive and the positivity was GM dose dependent. Therefore, this functional recovery of transduction activity was not owing to the proliferation of hair cells but was likely the self-repair of the hair bundle.

Innervation of stem cell-derived neurons into auditory epithelia of mice.

This study examined the potential of embryonic stem cell-derived neurons as transplants for cell therapy for the primary loss of spiral ganglion neurons. To assess the ability of embryonic stem cell-derived neurons for innervation into auditory epithelia, they were cocultured with auditory epithelium explants of mice for 7 days. Histological analysis demonstrated massive elongation of neurites from embryonic stem cell-derived neurons toward auditory hair cells. Embryonic stem cell-derived neurites were adjacent to or surrounding hair cells, and exhibited expression of synaptophysin, a marker for synaptic vesicles. These findings demonstrate the ability of embryonic stem cell-derived neurons for reinnervation into auditory epithelia, indicating a high potential of embryonic stem cell-derived neurons as transplants for replacing spiral ganglion neurons.

Nuclear translocation of beta-catenin in developing auditory epithelia of mice.

Beta-catenin, a protein component of adherens junctions, plays a role in the signalling pathway for cell proliferation. In this study, we examined the cellular distribution of beta-catenin in developing auditory epithelia of mice. Immunohistochemistry for Ki67 and cyclin D indicated active cell proliferation in premature auditory epithelia. In this period, the nuclear localization of beta-catenin in epithelial cells was observed together with expression of the lymphoid enhancer factor, a transcription factor in beta-catenin signalling. Epithelial cells showing nuclear localization of beta-catenin disappeared at the same time, as there was a decrease of cell proliferation. These findings indicate that nuclear translocation of beta-catenin plays a role in cell proliferation in developing auditory epithelia.

Electron transfer mediator micro-biosensor fabrication by organic plasma process.

We propose a new strategy for constructing a mediator-type biosensor as a Bio-MicroElectroMechanical Systems (BioMEMS) application. A vinylferrocene plasma-polymerized film (PPF) was deposited directly onto the surface of an electrode under dry conditions. The resulting redox film was extremely thin, adhered well onto a substrate (electrode), and had a highly crosslinked network structure. This technique, capable of polymeric deposition of any kind of monomer, can also serve the purpose of anti-fouling coating, or layer-to-layer interface creation. With a subsequent plasma process, additional polymeric layer of hydrophilic acetonitrile was superimposed onto the existing vinylferrocene-PPF surface to offer crucial features such that the wettability could be adjusted for a better electron transfer, and amino functional groups could be attached to immobilize a large amount of enzyme. Based upon this scheme, the device fabrication could be designed in a manner that the whole procedure was made up of dry wafer-handling processes, which is compatible with mass production. A prototype device was fabricated to have an array of needle-shaped amperometric micro-biosensors. The resultant thin polymer layer carried a large number of the mediator molecules, accomplishing a lower overpotential (+410 mV) and a rapid response time (<5s). Stressing the advantages of the plasma polymerization process together with some additional features accomplished in our device fabrication, we would discuss new possibilities in the field of BioMEMS.

Neural connections between embryonic stem cell-derived neurons and vestibular hair cells in vitro.

This study aimed to examine the potential of embryonic stem cell (ESC)-derived neural progenitors for restoration of the neural network in the peripheral vestibular system. Mouse ESC-derived neural progenitors were co-cultured with explants of vestibular sensory epithelia from neonatal mice. Histological analyses demonstrated that ESC-derived neurons substantially elongated their neurites towards vestibular hair cells, and attached to hair cells at the regions corresponding to the location of nerve endings in normal vestibular epithelia. Immunoreactivity for synaptophysin, a marker for synaptic vesicles, was present only in the cytoplasm of hair cells in sensory epithelia cultured alone, while the nerve endings of ESC-derived neurons attached to hair cells exhibited intense immunoreactivity for synaptophysin and some hair cells were moderately reactive in co-cultured specimens. The pattern of synaptophysin expression in co-cultured specimens was very similar to that observed in developing sensory epithelia, in which synaptic connections between hair cells and nerve endings are actively formed. These findings indicate that ESC-derived neurons have the potential to restore neural connections in the peripheral vestibular system.

Rat neurological disease creeping is caused by a mutation in the reelin gene.

Reelin (Reln) is an extracellular matrix protein secreted from distinct neuronal populations and controls neural cell positioning during brain development. Alterations of human RELN have been reported in two pedigrees with an autosomal recessive lissencephaly. Although several alleles of the mouse reeler mutation were identified as disruptions of Reln, there is no other animal model with a confirmed mutation in Reln. We recently established the Komeda Zucker creeping (KZC) rat strain with an autosomal recessive mutation creeping (cre), showing a reeler-like phenotype. We also found that creeping was located in the genomic segment on rat chromosome 4 containing Reln and that the expression level of Reln mRNA was markedly reduced in cre/cre homozygous mutant animals. Here we report positional candidate cloning of creeping, and identify a nucleotide insertion mutation in Reln. This mutation leads to a translational frameshift and results in truncation of the predicted protein in the fourth reelin-specific repeat, removing the C-terminal region required for secretion and function of the protein. We conclude that the mutation detected here is causative and is probably a null allele. The KZC rat is the first rat model with a confirmed Reln mutation and would therefore contribute to the understanding of the Reln function.

Arrest of apoptosis in auditory neurons: implications for sensorineural preservation in cochlear implantation.

HYPOTHESIS: The JNK/c-Jun cell death pathway is a major pathway responsible for the loss of oxidative stress-damaged auditory neurons. BACKGROUND: Implantation of patients with residual hearing accentuates the need to preserve functioning sensorineural elements. Although some auditory function may survive electrode insertion, the probability of initiating an ongoing loss of auditory neurons and hair cells is unknown. Cochlear implantation can potentially generate oxidative stress, which can initiate the cell death of both auditory neurons and hair cells. METHODS: Dissociated cell cultures of P4 rat auditory neurons identified the apoptotic pathway initiated by oxidative stress insults (e.g., loss of trophic factor support) and characterized this pathway by arresting translation of pathway-specific mRNA with antisense oligonucleotide treatment and with the use of pathway specific inhibitors. The presence or absence of apoptosis-specific protein and changes in the level of neuronal survival measured the efficacy of these interventional strategies. RESULTS: These in vitro studies identified the JNK/c-Jun cascade as a major initiator of apoptosis of auditory neurons in response to oxidative stress. Neurons pretreated with c-jun antisense oligonucleotide and exposed to high levels of oxidative stress were rescued from apoptosis, whereas neurons in treatment control cultures died. Treatment of oxidative-stressed cultures with either curcumin, a MAPKKK pathway inhibitor, or PD-098059, a MEK1 inhibitor, blocked loss of neurons via the JNK/c-Jun apoptotic pathway. CONCLUSION: Blocking the JNK/c-Jun cell death pathway is a feasible approach to treating oxidative stress-induced apoptosis within the cochlea and may have application as an otoprotective strategy during cochlear implantation.

Evidence for tachykinin NK3 receptors-triggered peptide YY release from isolated guinea-pig distal colon.

The anorectic gut hormone, peptide YY (PYY), is released from colonic mucosal endocrine cells, but little is known about the role for tachykinin NK3 receptor in the control of PYY release from the colonic mucosa. We investigated the functional role for NK3 receptors in the control of PYY release from isolated guinea-pig distal colon, and the role for NK3 receptors-triggered PYY release in the control of colonic motility. Isolated colonic preparations were mounted in organ baths for measurement of PYY release and mechanical activity. The release of PYY from these preparations was determined by enzyme immunoassays. The NK3 receptor agonist senktide produced a tetrodotoxin/atropine-sensitive sustained increase in the release of PYY from the colonic preparations. Basal PYY release was transiently inhibited by the NK3 receptor antagonist SB222200. The neuropeptide Y1 receptor antagonist BIBO3304 produced a leftward shift of the concentration-response curves for senktide-evoked neurogenic contraction, but neither the neuropeptide Y2 receptor antagonist BIIE0246 nor the neuropeptide Y5 receptor antagonist CGP71683 affected the senktide concentration-response curves. NK3 receptors appear to play an important role in the control of PYY release from colonic mucosa, and NK3 receptor-triggered PYY release can exert Y1 receptor-mediated inhibition of tachykinergic neuromuscular transmission. This indicates a pathophysiological role for the NK3 receptor-triggered PYY release in the control of colonic motility.

Intranasal immunization with a non-adjuvanted adhesive protein descended from Pasteurella pneumotropica and its preventive efficacy against opportunistic infection in mice.

Intranasal vaccination is one of the most effective means of protecting against invading and colonizing pathogens because the vaccine elicits a mucosal immune response. The exploitation of vaccine adjuvants and delivery systems for intranasal vaccines is an important way to evoke antigen immunogenicity and elicit a better immune response at the mucosal sites. In the present study, we assessed the potential of intranasal immunization using a non-adjuvanted bacterial adhesive protein toward the host organs. We evaluated intranasal immunization with modified recombinant PnxIIIA (MP3) from Pasteurella pneumotropica and its preventive efficacy against opportunistic infection caused by P. pneumotropica, without using any adjuvants or delivery systems. The 100-kDa MP3 was confirmed to retain its immunogenicity and binding activity to collagen type I similar to the parent PnxIIIA. When MP3 was fused to green-fluorescent protein and inoculated into C57BL/6J mice intranasally, fluorescence intensity in the intranasal airway could be observed until 3 h after inoculation. Mice were intranasally immunized with MP3 at a maximum of 4 doses, with 7-day intervals. The antibody titer of serum IgG and IgA specific for MP3, as well as that of bronchoalveolar lavage fluid IgA, showed more than 9 (log2) after 3 or 4 rounds of immunization. Experimentally infecting immunized mice with P. pneumotropica resulted in the inability to isolate the bacterium from the nasal cavity, trachea, conjunctiva, or cecum with more than 3 doses in the immunized mice. Although the detection in each organ seldom changed with less than 2 rounds of immunization, unlike that observed in the non-immunized mice, the detection remarkably decreased with 3 or more rounds of immunization. These results suggest that intranasal immunization with a non-adjuvanted adhesive protein could have preventive effects against opportunistic infection by P. pneumotropica.

Mechanical stress-induced reactive gliosis in the auditory nerve and cochlear nucleus.

OBJECT: Hearing levels following microsurgical treatment gradually deteriorate in a number of patients treated for vestibular schwannoma (VS), especially in the subacute postoperative stage. The cause of this late-onset deterioration of hearing is not completely understood. The aim of this study was to investigate the possibility that reactive gliosis is a contributory factor. METHODS: Mechanical damage to nerve tissue is a feature of complex surgical procedures. To explore this aspect of VS treatment, the authors compressed rat auditory nerves with 2 different degrees of injury while monitoring the compound action potentials of the auditory nerve and the auditory brainstem responses. In this experimental model, the axons of the auditory nerve were quantitatively and highly selectively damaged in the cerebellopontine angle without permanent compromise of the blood supply to the cochlea. The temporal bones were processed for immunohistochemical analysis at 1 week and at 8 weeks after compression. RESULTS: Reactive gliosis was induced not only in the auditory nerve but also in the cochlear nucleus following mechanical trauma in which the general shape of the auditory brainstem response was maintained. There was a substantial outgrowth of astrocytic processes from the transitional zone into the peripheral portion of the auditory nerve, leading to an invasion of dense gliotic tissue in the auditory nerve. The elongated astrocytic processes ran in parallel with the residual auditory neurons and entered much further into the cochlea. Confocal images disclosed fragments of neurons scattered in the gliotic tissue. In the cochlear nucleus, hypertrophic astrocytic processes were abundant around the soma of the neurons. The transverse diameter of the auditory nerve at and proximal to the compression site was considerably reduced, indicating atrophy, especially in rats in which the auditory nerve was profoundly compressed. CONCLUSIONS: The authors found for the first time that mechanical stress to the auditory nerve causes substantial reactive gliosis in both the peripheral and central auditory pathways within 1-8 weeks. Progressive reactive gliosis following surgical stress may cause dysfunction in the auditory pathways and may be a primary cause of progressive hearing loss following microsurgical treatment for VS.

Selective vulnerability of adult cochlear nucleus neurons to de-afferentation by mechanical compression.

It is well established that the cochlear nucleus (CN) of developing species is susceptible to loss of synaptic connections from the auditory periphery. Less information is known about how de-afferentation affects the adult auditory system. We investigated the effects of de-afferentation to the adult CN by mechanical compression. This experimental model is quantifiable and highly reproducible. Five weeks after mechanical compression to the axons of the auditory neurons, the total number of neurons in the CN was evaluated using un-biased stereological methods. A region-specific degeneration of neurons in the dorsal cochlear nucleus (DCN) and posteroventral cochlear nucleus (PVCN) by 50% was found. Degeneration of neurons in the anteroventral cochlear nucleus (AVCN) was not found. An imbalance between excitatory and inhibitory synaptic transmission after de-afferentation may have played a crucial role in the development of neuronal cell demise in the CN. The occurrence of a region-specific loss of adult CN neurons illustrates the importance of evaluating all regions of the CN to investigate the effects of de-afferentation. Thus, this experimental model may be promising to obtain not only the basic knowledge on auditory nerve/CN degeneration but also the information relevant to the application of cochlear or auditory brainstem implants.

An animal experimental model of auditory neuropathy induced in rats by auditory nerve compression.

Several animal models of auditory neuropathy (AN) have been produced by employing pharmacological agents to damage auditory neurons or hair cells selectively. The specificity of pharmacological lesions is generally assessed by observation of visible structural damage but it is difficult to localize the delivery, which could lead to functional side effects in other anatomical structures. Although genetic analyses of human AN patients have provided important information on the pathophysiology of AN, specific genetic defects have not been fully correlated with functional deficits in the auditory nervous system. To address this problem, we compressed rat auditory nerves to assess neural degeneration for up to 35 weeks. The method produced a good model of auditory neuropathy, including profound deterioration of the auditory brainstem response and preservation of both cochlear microphonics and distortion product otoacoustic emissions. Histological examination revealed that in spite of profound degeneration of the auditory nerve, the hair cells remained intact. The model provides a complementary alternative to those based on pharmacological lesions and genetic analyses of AN patients and should allow analysis of the pathophysiology of auditory neuropathy with less risk of the results being confounded by unknown deficits in other cell types.