Phantom auditory perception (tinnitus): mechanisms of generation and perception

Normal hearing depends on sound amplification within the mammalian cochlea. The amplification, without which the auditory system is effectively deaf, can be traced to the correct functioning of a group of motile sensory hair cells, the outer hair cells of the cochlea. Acting like motor cells, outer hair cells produce forces that are driven by graded changes in membrane potential. The forces depend on the presence of a motor protein in the lateral membrane of the cells. This protein, known as prestin, is a member of a transporter superfamily SLC26. The functional and structural properties of prestin are described in this review. Whether outer hair cell motility might account for sound amplification at all frequencies is also a critical question and is reviewed here.

Cochlear Outer Hair Cell Motility

Aminoglycosides are commonly prescribed antibiotics with deleterious side effects to the inner ear. Due to their popular application as a result of their potent antimicrobial activities, many efforts have been undertaken to prevent aminoglycoside ototoxicity. Over the years, understanding of the antimicrobial as well as ototoxic mechanisms of aminoglycosides has increased. These mechanisms are reviewed in regard to established and potential future targets of hair cell protection.

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Mechanisms of Aminoglycoside Ototoxicity and Targets of Hair Cell Protection

The most serious side-effect of the widely used aminoglycoside antibiotics is irreversible intracellular damage to the auditory and vestibular hair cells of the inner ear. The mechanism of entry into the hair cells has not been unequivocally resolved. Here we report that extracellular dihydrostreptomycin not only blocks the mechano-electrical transducer channels of mouse outer hair cells at negative membrane potentials, as previously shown, but also enters the cells through these channels, which are located in the cells' mechanosensory hair bundles. The voltage-dependent blocking kinetics indicate an open-channel block mechanism, which can be well described by a two barrier–one binding site model, quantifying the antibiotic's block of the channel as well as its permeation in terms of the associated rate constants. The results identify the open transducer channels as the main route for aminoglycoside entry. Intracellularly applied dihydrostreptomycin also blocks the transducer channels, but at positive membrane potentials. However, the potency of the block was two orders of magnitude lower than that due to extracellular dihydrostreptomycin. Extracellular Ca2+ increases the free energy of the barrier nearest the extracellular side and of the binding site for dihydrostreptomycin. This reduces both the entry of dihydrostreptomycin into the channel and the channel's affinity for the drug. In vivo, where the extracellular Ca2+ concentration in the endolymph surrounding the hair bundles is < 100 μm, we predict that some 9000 dihydrostreptomycin molecules per second enter each hair cell at therapeutic drug concentrations.

https://physoc.onlinelibrary.wiley.com/doi/pdf/10.1113/jphysiol.2005.085951

The aminoglycoside antibiotic dihydrostreptomycin rapidly enters mouse outer hair cells through the mechano-electrical transducer channels.

Recordings were made from single afferent fibers in the inferior vestibular nerve, which innervates the saccule and posterior semicircular canal. A substantial portion of the fibers with irregular background activity increased their firing in response to moderately intense clicks and tones. In responsive fibers, acoustic clicks evoked action potentials with minimum latencies of 80 dB SPL) caused synchronization of spikes to preferred phases of the tone cycle. PUSH and PULL fibers had preferred response phases approximately 180 degrees apart. These two response classes are consistent with fibers that innervate hair cells having opposite morphological polarizations, an arrangement found in the saccule. With low-frequency tone bursts, sound levels of > or = 90 dB SPL evoked increases in mean spike rate. Spike rates increased monotonically with sound level without saturating at levels < or = 115 dB SPL. Contraction of the middle-ear muscles decreased responses to sound, consistent with the sound transmission path being through the middle ear. Several fibers were labeled with biocytin and traced. All labeled fibers had bipolar cell bodies in the inferior vestibular ganglion with peripheral processes extending toward the saccular nerve and central processes in the vestibular nerve. Two fibers were traced to the saccular epithelium. One fiber was traced centrally and arborized extensively in vestibular nuclei and a region ventromedial to the cochlear nucleus. Our results confirm and extend previous suggestions that the mammalian saccule responds to sound at frequencies and levels within the normal range of human hearing. We suggest a number of auditory roles that these fibers may play in the everyday life of mammals.

https://www.jneurosci.org/content/jneuro/14/10/6058.full.pdf

Acoustically responsive fibers in the vestibular nerve of the cat

Cochlear outer hair cells have been well established as primary targets of the ototoxic actions of aminoglycoside antibiotics. These cells, isolated from the guinea pig cochlea and maintained in short-term culture, were used as a model for evaluating the acute effects of gentamicin on cell viability, depolarization-induced transmembrane calcium flux, and depolarization-induced motile responses.



On the basis of morphology and fluorochromasia, the presence of extracellular gentamicin as high as 5 mM did not affect the viability of the cells for up to 6 hr, the longest time tested. Viable cells showed binding of fluorescently tagged gentamicin to their base but excluded the drug from their cytoplasm. In response to [K+]-depolarization, intracellular calcium levels (monitored with the fluorescent calcium-sensitive dye fluo-3) increased from a resting value of 218 ± 102 nM to 2,018 ± 1,077 nM concomitant with a cell shortening of 0.7% ± 1.3%. The depolarization-induced calcium increase was apparently caused by calcium entry into the cell as it was inhibited by the calcium-channel blocker methoxyverapamil and prevented in the absence of extracellular calcium. Both gentamicin and neomycin blocked the [K+]-induced calcium increase at an IC50 of 50 μM. Despite the inhibition of calcium entry the ability of the outer hair cells to shorten under [K+]-depolarization was not impaired; in fact, cell shortening was even more pronounced in the absence of calcium influx (2.6% ± 1.4%). This argues effectively against the existence of a calcium-dependent actomyosin-mediated component in [K+]-induced shape changes.



The results suggest the existence of voltage-gated calcium channels in outer hair cells and that calcium influx through these channels is impaired by the aminoglycoside antibiotics neomycin and gentamicin. This action may be part of the acute ototoxic mechanism of these molecules. Furthermore, the results not only confirm the calcium independence of the depolarization-induced motility but also suggest that calcium influx into outer hair cells opposes cell shortening.

/pdf/aminoglycoside-antibiotics-impair-calcium-entry-but-not-2ocglmfptb.pdf

Aminoglycoside antibiotics impair calcium entry but not viability and motility in isolated cochlear outer hair cells.

Potassium-depolarization induces motility in isolated outer hair cells by an osmotic mechanism

Measures of cochlear selectivity can be obtained from compound responses using tone‐on‐tone masking procedures [Dallos and Cheatham, J. Acoust. Soc. Am. 59, 591–597 (1976)]. For the normal guinea pig, cochlear fiber tuning is sharper by a factor of 1.8 than AP tuning curves using simultaneous masking (threshold criterion  = 25% N1 amplitude reduction). Anesthesia does not appear to affect AP tuning. In pathological cochleas, AP tuning is broadened by a factor of 2–3, and differences between forward and simultaneous masking curves are reduced. Tuning changes can sometimes occur without threshold elevation. AP tuning curves were obtained from humans during transtympanic electrocochleography. For subjects with near normal thresholds, Q10 dB values (simultaneous masking) are approximately 2.3 at 2 kHz, 3.6 at 4 kHz, and 4.7 at 8 kHz. Using the relationship between cochlear fiber tuning and AP tuning in the guinea pig, estimates of human cochlear fiber tuning are 4.2 at 2 kHz, 6.5 at 4 kHz, and 8.5 at 8 kHz. Patients with threshold elevations of more than 30 dB resulting from cochlear deafness have AP tuning curves less sharply tuned by a factor of 2–3.

AP tuning curves from normal and pathological human and guinea pig cochleas

Large-conductance calcium-activated potassium (BK) channels are known to play a prominent role in the hair cell function of lower vertebrates where these channels determine electrical tuning and re...

Contribution of BK Ca2+-Activated K+ Channels to Auditory Neurotransmission in the Guinea Pig Cochlea

Immunodetection of gentamicin (GM) was carried out on surface preparations of the whole organ of Corti from cochleas of guinea pigs treated daily with GM at a dose of 60 mg/kg/day and sacrificed at the end of different treatment periods. Cochlear function was determined just before sacrifice, 24 h after the last injection. Threshold elevations, mainly at high frequencies, were noted only after 10-14 days of treatment. However, the presence of GM was observed much earlier, as early as after the second injection, and specifically in the sensory hair cells. GM labelling was essentially observed in the outer hair cells (OHC) and increased from the apex to the base of the cochlea and from the third to the first row of OHC. GM labelling of inner hair cells was less pronounced and was observed only after the 8th day of treatment. These observations demonstrate that GM specifically enters and accumulates in the sensory hair cells and that the uptake precedes the development of functional and cellular damage which may result from a long-term intracellular cytotoxic action of the molecule.

Jean Marie Aran

Papers

Aminoglycoside antibiotics impair calcium entry but not viability and motility in isolated cochlear outer hair cells.

Potassium-depolarization induces motility in isolated outer hair cells by an osmotic mechanism

AP tuning curves from normal and pathological human and guinea pig cochleas

Contribution of BK Ca2+-Activated K+ Channels to Auditory Neurotransmission in the Guinea Pig Cochlea

Gentamicin Uptake by Cochlear Hair Cells Precedes Hearing Impairment during Chronic Treatment