C-Phycocyanin Attenuates Noise-Induced Cochlear Synaptopathy via the Inhibition of Oxidative Stress and Intercellular Adhesion Molecule-1 in the Cochlea.

The synapses between inner hair cells (IHCs) and spiral ganglion neurons (SGNs) are the most vulnerable structures in the noise-exposed cochlea. Cochlear synaptopathy results from the disruption of these synapses following noise exposure and is considered the main cause of poor speech understanding in noisy environments, even when audiogram results are normal. Cochlear synaptopathy leads to the degeneration of SGNs if damaged IHC-SGN synapses are not promptly recovered. Oxidative stress plays a central role in the pathogenesis of cochlear synaptopathy. C-Phycocyanin (C-PC) has antioxidant and anti-inflammatory activities and is widely utilized in the food and drug industry. However, the effect of the C-PC on noise-induced cochlear damage is unknown. We first investigated the therapeutic effect of C-PC on noise-induced cochlear synaptopathy. In vitro experiments revealed that C-PC reduced the H2O2-induced generation of reactive oxygen species in HEI-OC1 auditory cells. H2O2-induced cytotoxicity in HEI-OC1 cells was reduced with C-PC treatment. After white noise exposure for 3 h at a sound pressure of 118 dB, the guinea pigs intratympanically administered 5 µg/mL C-PC exhibited greater wave I amplitudes in the auditory brainstem response, more IHC synaptic ribbons and more IHC-SGN synapses according to microscopic analysis than the saline-treated guinea pigs. Furthermore, the group treated with C-PC had less intense 4-hydroxynonenal and intercellular adhesion molecule-1 staining in the cochlea compared with the saline group. Our results suggest that C-PC improves cochlear synaptopathy by inhibiting noise-induced oxidative stress and the inflammatory response in the cochlea.

Assessment of the expression and role of the α1-nAChR subunit in efferent cholinergic function during the development of the mammalian cochlea.

Hair cell (HC) activity in the mammalian cochlea is modulated by cholinergic efferent inputs from the brainstem. These inhibitory inputs are mediated by calcium-permeable nicotinic acetylcholine receptors (nAChRs) containing α9- and α10-subunits and by subsequent activation of calcium-dependent potassium channels. Intriguingly, mRNAs of α1- and γ-nAChRs, subunits of the "muscle-type" nAChR have also been found in developing HCs (Cai T, Jen HI, Kang H, Klisch TJ, Zoghbi HY, Groves AK. J Neurosci 35: 5870-5883, 2015; Scheffer D, Sage C, Plazas PV, Huang M, Wedemeyer C, Zhang DS, Chen ZY, Elgoyhen AB, Corey DP, Pingault V. J Neurochem 103: 2651-2664, 2007; Sinkkonen ST, Chai R, Jan TA, Hartman BH, Laske RD, Gahlen F, Sinkkonen W, Cheng AG, Oshima K, Heller S. Sci Rep 1: 26, 2011) prompting proposals that another type of nAChR is present and may be critical during early synaptic development. Mouse genetics, histochemistry, pharmacology, and whole cell recording approaches were combined to test the role of α1-nAChR subunit in HC efferent synapse formation and cholinergic function. The onset of α1-mRNA expression in mouse HCs was found to coincide with the onset of the ACh response and efferent synaptic function. However, in mouse inner hair cells (IHCs) no response to the muscle-type nAChR agonists (±)-anatoxin A, (±)-epibatidine, (-)-nicotine, or 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP) was detected, arguing against the presence of an independent functional α1-containing muscle-type nAChR in IHCs. In α1-deficient mice, no obvious change of IHC efferent innervation was detected at embryonic day 18, contrary to the hyperinnervation observed at the neuromuscular junction. Additionally, ACh response and efferent synaptic activity were detectable in α1-deficient IHCs, suggesting that α1 is not necessary for assembly and membrane targeting of nAChRs or for efferent synapse formation in IHCs.

Challenges and opportunities in presbycusis.

The population aged 65 years and older is increasing at a faster rate than the total population, with predictions that by 2030, 20% of the population will be 65 years or older. In 2006, between 35% and 50% of those aged 65 years or older reportedly had presbycusis, a sensory impairment that contributes to social isolation and loss of autonomy and is associated with anxiety, depression, and cognitive decline. To address these concerns, the Geriatric Committee of the American Academy of Otolaryngology, in conjunction with the Hearing Committee, focused on 3 challenges and opportunities in the management of presbycusis: (1) the financial burden of caring for patients with presbycusis in the face of increasing costs and declining reimbursements; (2) future treatment options arising from improved understanding of the molecular mechanisms underlying presbycusis, and (3) recognition of central presbycusis as a condition commonly superimposed on peripheral age-related hearing loss whose diagnosis and management can improve outcomes.

Experimental autoimmune labyrinthitis: assessment of molecular size of autoantigens in fractions of inner ear proteins eluted on the Mini Whole Gel Eluter.

In order to determine the molecular size of the causative autoantigens of experimental autoimmune labyrinthitis, crude inner ear antigen was separated into 14 fractions and the constituent molecules were identified by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis using the Mini Whole Gel Eluter. The mean total protein recovery was approximately 66%. Sensitization with 55-65 kDa proteins induced the highest number of infiltrating cells among the fractions and sensitization with 38-45 kDa proteins induced the second highest number of inflammatory cells. These results suggest that 38-45 kDa and 55-65 kDa proteins are the causative autoantigens of experimental autoimmune labyrinthitis.

[3H]-aldosterone binding sites (type I receptors) in the lateral wall of the cochlea: distribution assessment by quantitative autoradiography.

Mineralocorticoids, such as aldosterone, are steroids that enhance Na+ retention and K+ excretion in ion-transporting epithelial tissues through regulation of Na, K-ATPase. Previous studies suggest that aldosterone may regulate labyrinthine ion transport through up-regulation of Na, K-ATPase sites, a process mediated by high-affinity aldosterone (type I) receptors. In the present study, information concerning density and distribution of aldosterone binding sites in cochlear lateral wall tissues was determined by quantitative autoradiography using [3H]-aldosterone and RU-28362, a glucocorticoid agonist that blocks low affinity binding to glucocorticoid (type II) receptors. The results revealed that the distribution of aldosterone binding sites differs among the individual cochlear regions of the lateral wall. The highest level of binding was associated with the stria vascularis and epithelial cells of the spiral prominence. Elevated levels of binding were also observed in stromal cells of the spiral prominence, and to a lesser extent in the spiral ligament. The differential distribution of aldosterone binding sites in the lateral wall resembles the pattern of localization of Na, K-ATPase sites observed in previous studies and is compatible with the idea that mineralocorticoids play a role in the regulation of cochlear cation transport.

Quantitative assessment of perilymph sources.

The problem of the perilymph origin--influx of cerebrospinal fluid (CSF) versus ultrafiltration within the cochlea--cannot be solved by mere qualitative proofs of tracer passage through the cochlear aqueduct. In order to gain quantitative data on the possible perilymph sources, an experimental study was designed to follow the time course of dye concentrations in the cisternal CSF and in the perilymph after tracer injection into the CSF at the vertex. By comparing the resulting concentration peaks in both fluids, the mean peak of the perilymph tracer concentrations was found to reach 36% of the maximum CSF concentration only. It is concluded that the local perilymph production within the cochlea exceeds the influx of CSF by a ratio of about 2:1. A working hypothesis of the double perilymph origin is discussed.