Biomedical subjects

L D Fechter

Publications and source records attributed to L D Fechter.

At least 19 recordsLinked to original sources

PubMed2001

NMDA receptor blockage protects against permanent noise-induced hearing loss but not its potentiation by carbon monoxide.

While a clear role has been proposed for glutamate as a putative neurotransmitter at the inner hair cell type I spiral ganglion cell synapse, the possible role of excessive glutamate release in cochlear impairment and of NMDA receptors in such a process is uncertain. The present study compares the protective effects of (+)-MK-801, an NMDA receptor antagonist, and the relatively inactive isomer (-)-MK-801 against permanent noise-induced hearing loss (NIHL). The study also asks whether (+)-MK-801 can protect against the NIHL potentiation by carbon monoxide (CO). Rats (n = 6) were exposed to 100-dB, 13.6-kHz octave-band noise for 2 h after receiving injection of (+)-MK-801 hydrogen maleate (1 mg/kg), (-)-MK-801 hydrogen maleate (1 mg/kg), or saline. Other groups of animals were exposed to the combination of noise and CO (1200 ppm) after receiving (+)-MK-801 or saline. Additional subjects received (+)-MK-801, saline or CO exposure alone. Compound action potential (CAP) threshold sensitivities were compared 4 weeks after the exposures. The results show significant protection by (+)-MK-801 against the permanent CAP threshold elevation induced by noise alone, but no protective effect of (-)-MK-801. (+)-MK-801 produced limited protection against threshold shifts induced by the combination of noise and CO. Outer hair cell (OHC) loss was not protected by (+)-MK-801 administration. The data suggest that NMDA receptor stimulation may play a role in NIHL resulting from fairly mild noise exposure. The data do not support a role for NMDA receptor stimulation in the potentiation of NIHL that results from simultaneous exposure to CO and noise.

Action Potentials↗

PubMed2001

Free radical generation in the cochlea during combined exposure to noise and carbon monoxide: an electrophysiological and an EPR study.

Ototoxicity following combined exposure to noise and carbon monoxide (CO) is known to result in more severe permanent threshold shifts than exposure to noise alone. We have previously demonstrated that such potentiation of noise-induced auditory impairment by CO can be prevented by the administration of a nitrone spin-trapping agent. Although such protection implicates injury via free radical pathways, drug-induced protection does not provide direct evidence for the presence of free radicals in the cochlea. The objective of this study was to demonstrate the actual presence of nitrone spin adducts in the cochlea following simultaneous exposure to noise and CO. Using electrophysiological end-points, the protective effects of the nitrone spin-trapping agent alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone (POBN) were assessed following combined exposure of adult male Long Evans hooded rats to noise and CO. In addition, an ex-vivo evaluation of POBN spin adducts was done by electron paramagnetic resonance spectroscopy (EPR). The noise used was octave band noise with center frequency 13.6 kHz at 100 dB(Lin) for a duration of 2 h. The level of CO used was 1200 ppm. Electrophysiological results demonstrate that POBN protects against combined exposure to noise plus CO. The EPR study demonstrates POBN spin adducts in the cochleae of animals exposed to noise plus CO. Therefore, this study provides evidence to the hypothesis that ototoxicity due to noise plus CO exposure is mediated via free radicals.

Animals↗

PubMed2000

Protective effects of phenyl-N-tert-butylnitrone on the potentiation of noise-induced hearing loss by carbon monoxide.

Free radical injury has been implicated in cochlear damage resulting from exposure to high-intensity noise and due to carbon monoxide (CO) hypoxia. Although exposure to noise plus CO is common in occupational settings and noise-induced hearing loss (NIHL) is enhanced in the presence of CO, potential mechanisms resulting in auditory impairment have not been studied. This study evaluates protective effects of the free radical scavenger phenyl-N-tert-butylnitrone (PBN) against potentiation of NIHL by CO. Three PBN administration protocols have been evaluated in subjects exposed to noise plus CO or noise alone. Long Evans hooded rats were exposed to octave band noise at 100 dB(Lin), center frequency (cf) = 13.6 kHz for a duration of 2 h. The level of CO used was 1200 ppm. Endpoints used to detect permanent auditory impairment were compound action potential (CAP) threshold and 1 microV root mean square (RMS) cochlear microphonic (CM). Testing was done 4 weeks following exposure. PBN administration prior to and following simultaneous exposure provided significant protection against auditory impairment in subjects receiving noise plus CO. Partial protection was observed in the protocols where PBN was injected following noise plus CO exposure. PBN administration appeared to reduce auditory impairment in animals exposed to noise alone, but the difference was not found to be statistically significant. Protective effects of PBN following simultaneous exposure to noise plus CO suggest that free radicals may be generated during combined exposure.

Animals↗

PubMed2000

Low-level toluene disrupts auditory function in guinea pigs.

Toluene appears to have adverse effects on the human auditory system, but it is difficult to estimate its potency since it is commonly present in the workplace in combination with noise exposure; workplace noise exposures are often highly variable. Studies designed to assess toluene ototoxicity specifically have been limited to high-dose studies in a single laboratory animal model, the rat. Here permanent hearing loss has been observed at concentrations of 1000 ppm toluene and greater after inhalation exposure for 5 days, 6 h/day. The OSHA threshold limit value for toluene is only 100 ppm. The current study focuses on the onset of toluene ototoxicity acutely in the guinea pig and in adducing a mechanism of effect. In this study, evidence is presented for the impairment of auditory function by toluene in the guinea pig, at a concentration substantially lower than that used for studying permanent impairment in the rat. The impaired function was correlated with reduced energy metabolism in outer hair cells. Assessment of auditory function was made using distortion product otoacoustic emissions (DPOAE) with subsequent measurement of succinate dehydrogenase (SDH) staining density in hair cells using surface preparations. Temporary disruption of auditory function in guinea pigs is seen in subjects exposed to 250, 500, and 1000 ppm toluene for 8 h/day, 5 day/week for 1 and 4 weeks. Concentrations as low as 250 ppm toluene were able to disrupt auditory function acutely in the guinea pig, and 500 and 1000 ppm toluene produced greater acute dysfunction. SDH staining suggests that reduced enzyme activity in the midfrequency region of the cochlea occurs acutely following toluene exposure. Although the auditory dysfunction progressed between 1 and 4 weeks of exposure, a permanent loss did not develop for these subjects and hair cell death was not seen. The current study identifies early evidence of auditory system impairment in the guinea pig at low toluene concentration and evidence for impairment of energy production in hair cells. While even a transient auditory impairment has implications for workplace safety, additional study on the transition from such acute effects to permanent impairment is essential.

Animals↗

PubMed2000

Succinate dehydrogenase (SDH) activity in hair cells: a correlate for permanent threshold elevations.

Hair cell loss is often used as a histological correlate of hearing loss. However, the histological and the physiological data are not always well correlated. This paper investigates the use of succinate dehydrogenase (SDH) activity in the hair cells as a marker of cellular dysfunction and so the loss of auditory sensitivity. In our previous studies, potentiation of noise-induced auditory threshold elevation by carbon monoxide (CO) was observed [Chen and Fechter, 1999; Chen et al., 1999]. However, its histological basis is still unclear. In this study, rats were exposed to 100-dB octave-band noise (center frequency=13.6 kHz, 2 h) or to the combination of the noise and CO (1200 ppm). Threshold elevation of compound action potential (CAP) and cochlear histological changes were assessed 4 weeks after exposure. The noise alone caused CAP threshold elevations with little if any or without hair cell loss. However, the SDH activity in the hair cells decreased after the exposure. The SDH reduction, especially in the inner hair cells, was well related to the loss of auditory sensitivity. The combined exposure to noise and CO caused more severe CAP threshold elevation and SDH activity reduction than did the noise alone and it also caused significant outer hair cell loss. However, across all the test frequencies, neither the hair cell loss nor the SDH reduction alone had good correlation to the reduction of the auditory sensitivity. Under this situation, CAP threshold elevation seemed to follow OHC loss at high frequencies and to follow SDH reductions in the IHCs at low frequencies, where no hair cell loss occurred.

Action Potentials↗

PubMed2000

Simultaneous monitoring of slow cell motility and calcium signals of the guinea pig outer hair cells.

'Slow' motility (shape changes over seconds to minutes) of the mammalian cochlear outer hair cell (OHC) could play a protection role from intense sound pressure and is associated with elevation of the cytosolic free Ca(2+) concentration ([Ca(2+)](i)). In the present work, a new approach was elaborated using fluorescent imaging for continuous monitoring of both [Ca(2+)](i) changes and slow motility of OHCs employing the Ca(2+) fluorescent indicator Fura-2. Whole OHC fluorescence and that of cell segments were analyzed to discriminate between fluorescence changes caused by [Ca(2+)](i) rise and those related to change of the cell shape. The reliability of the method was examined by simultaneous monitoring of [Ca(2+)](i) and OHC length changes induced by change of buffer osmolarity or by increase of KCl concentration. The method revealed that the time course of [Ca(2+)](i) increase and rate of cell shortening often do not coincide. It was also observed that [Ca(2+)](i) increased in 70 mM KCl more slowly than the rate of KCl delivery to OHCs. The comparison of the time courses of [Ca(2+)](i) elevation, induced by increase of K(+)/Na(+) ratio and by substitution of Na(+) with N-methyl-D-glucamine(+), indicated that the relatively slow kinetics of [Ca(2+)](i) increase in the OHC is partially attributed to regulation of Ca(2+) homeostasis by the Na(+)/Ca(2+) exchanger.

Animals↗

PubMed2000

Increased noise severity limits potentiation of noise induced hearing loss by carbon monoxide.

This study evaluates the influence of noise intensity and duration on auditory dysfunction due to simultaneous exposure to noise and carbon monoxide (CO). Previous studies have demonstrated that CO potentiates noise induced hearing loss (NIHL). It is not known whether auditory dysfunction due to combined exposure parallels impairment due to noise alone. Based on the 5 dB exchange rate between noise intensity and exposure doubling time, equivalent noise exposure conditions were used. Long Evans hooded rats were divided into groups that received noise alone (95, 100 and 105 dB SPL), and noise plus CO (1200 ppm), for durations of 4, 2 and 1 h, respectively. Controls were exposed to air or CO alone. Thresholds were evaluated 4 weeks later using an electrophysiological endpoint, the compound action potential threshold. Results demonstrate that the 5 dB exchange rate is not conserved under the conditions and subjects used. Moreover, dysfunction due to combined exposure did not parallel dysfunction due to noise alone. Further, although an increase in exposure duration results in increased auditory dysfunction, no further potentiation of NIHL by CO is observed. This suggests that at increasing noise severity, dysfunction due to combined exposure is limited by impairment due to noise alone.

Acoustic Stimulation↗

PubMed2000

Characterization of the ototoxicity of difluoromethylornithine and its enantiomers.

Difluoromethylornithine (DFMO) is an irreversible inhibitor of ornithine decarboxylase (ODC), the essential enzyme in mammalian polyamine biosynthesis (Pasic et al., 1997, Arch. Otolaryngol. Head Neck Surg. 123[12], 1281-1286). This cancer chemotherapeutic agent has significant ototoxic potential. Because the DFMO enantiomers differ in their ability to block ODC, the present study was designed to compare the ototoxic potential of each enantiomer with the racemic form of this drug for the rat and guinea pig. Determining differential ototoxicity of the enantiomers is one preliminary step in determining the optimal form of DFMO to use in human cancer chemotherapy. Daily intubation with D,L-DFMO does not produce any auditory dysfunction in rats with doses between 200 mg/kg/day and 1. 2 g/kg/day for up to 8 weeks, despite the fact that doses of 800 and 1200 mg/kg/day depressed body weight gain. In contrast to the data observed in rats, substantial ototoxicity was observed when guinea pigs were injected ip with doses of D,L-DFMO between 500 mg/kg/day and 1 g/kg/day. D,L-DFMO produced loss of compound action potential sensitivity, but not of cochlear microphonic amplitude. This finding correlated with histological data revealing loss of both outer and inner hair cells in the cochlea with inner more affected than outer hair cells, particularly in the basal turn. Higher exposure doses (2-3 g/kg/day) resulted in significant general toxicity including impaired growth and some mortality. When the enantiomers were evaluated in the guinea pig, it was found that 1 g/kg/day D-DFMO did not produce any significant hearing impairment, whereas 1 g/kg/day of the L-enantiomer of DFMO generated a threshold shift that surpassed that of 1 g/kg/day of the D,L-DFMO treatment.

Animals↗

PubMed2000

Predicting exposure conditions that facilitate the potentiation of noise-induced hearing loss by carbon monoxide.

Hearing loss is the most common occupational disease in the United States, with noise serving as the presumed causative agent in most instances. This investigation characterizes the exposure conditions that facilitate the potentiation of noise-induced hearing loss (NIHL) by carbon monoxide (CO). Auditory function was compared in rats exposed 4 weeks earlier to noise alone, CO alone, combined exposure, and air in the exposure chamber. This interval between exposure and auditory threshold assessment was selected to permit recovery of temporary threshold shifts. The compound action potential (CAP) threshold evoked by pure tone stimuli was used as a measure of auditory sensitivity. The no adverse effect level (NOAEL) with respect to potentiation of NIHL was found to be 300 ppm CO. Potentiation of NIHL by CO increases linearly as CO concentration increases between 500 -1500 ppm. Benchmark dose software (version 1. 1B) published by the U.S. EPA National Center for Environmental Assessment was employed to determine a benchmark concentration of CO that produced either a 5-dB potentiation of NIHL or an increase in auditory threshold equivalent to 10% of the effect of noise alone. The lower bound for these benchmark concentrations were 320 and 194 ppm CO, respectively. Unlike CO dose, the relationship between noise severity and potentiation of NIHL by CO shows a nonlinear relationship. The greatest potentiation was observed at moderate noise exposures (100 dB, 2-h, octave band-limited noise, or OBN) that produce limited permanent threshold shifts. Repeated exposures to 95-dB noise for 2-h periods in combination with 1200 ppm CO also yielded potentiation of NIHL, though such effects were not observed following a single combined exposure. These results underscore the potential risk of hearing loss from combined exposure to noise and CO, and the risks associated with repeated exposure.

Action Potentials↗

PubMed1999

Potentiation of octave-band noise induced auditory impairment by carbon monoxide.

In previous studies from our lab, broadband noise induced hearing loss has been found to be potentiated by simultaneous carbon monoxide (CO) exposure. In the present study, octave-band noise induced auditory impairment was studied with the presence of CO at levels of 1500, 1200, 700, 500 and 300 ppm and zero (noise alone). Four octave-band noises (1.2-2.4, 2.4-4.8, 4.8-9.6 and 9.6-19.2 kHz) were used. Experimental subjects (rats) were grouped for the exposure (8 h) to each noise, CO and their combinations. The compound action potential (CAP) and cochlear microphonics (CM) were recorded 4 weeks after the exposure. The noise induced elevation of the CAP threshold and the CM iso-amplitude curve were potentiated by the simultaneous CO exposure when the CO level reached 500 ppm or higher. CO exposure alone had no effect on CAP or CM. The CO potentiation can occur in any frequency region depending on the noise band. The combined exposure can also induce threshold shifts in some cases in which both the noise and the CO alone did not cause threshold shifts. The size of the potentiation shown by CAP and CM was similar, indicating a possible origin of the CO potentiation from the damage to the outer hair cells. Interestingly, the hearing loss induced by noise alone gradually recovered (partially), but the hearing loss caused by the combined exposure did not. The potentiation may be due to the reduction of the cell's ability to repair noise induced damage by CO.

Animals↗

PubMed1999

Intermittent noise-induced hearing loss and the influence of carbon monoxide.

Intermittent noise causes less hearing loss than continuous noise of equal intensity. The reduction in damage observed with intermittent noise may be explained by the fact that the auditory system has time to recover between the noise phases. Simultaneous carbon monoxide (CO) exposure produces greater noise-induced hearing loss than does noise alone (Chen and Fechter, 1999). In the present study, intermittent noise (octave-band with a center frequency of 13.6 kHz, 100 dB) of a 2 h total duration but with a different duty cycle (% of noise during exposure) was used. The intermittent exposure that had a shorter noise duty cycle induced a less permanent threshold shift (PTS) than those that had a longer noise duty cycle (or less rest periods). This relation between the loss in compound action potential (CAP) sensitivity and the noise duty cycle (or rest period) was abolished by the presence of CO. The cochlear microphonic (CM) amplitude revealed similar results to those seen using the CAP. While intermittent noise that had a short noise duty cycle did not cause hair cell loss by itself, the combined exposure to noise and CO (1200 ppm) caused remarkable OHC loss in the basal turn.

Action Potentials↗

PubMed1998

Trichloroethylene ototoxicity: evidence for a cochlear origin.

Trichloroethylene (TCE) is known to produce an unusual pattern of hearing impairment in laboratory animals marked by a preferential loss of threshold sensitivity at midfrequencies. The purpose of this research was to determine whether the TCE-induced auditory deficit results from cochlear dysfunction. Adult Long Evans hooded rats were exposed via inhalation to either 0 (clean air) or 4000 ppm TCE (6 h/day for 5 days). Auditory thresholds for 1-40 kHz tones were determined 3 weeks after exposure using reflex modification audiometry (RMA; n = 12/group). Cochlear electropotentials were measured during subsequent testing (n = 3-10/group) 5 to 7 weeks after exposure, including thresholds for cochlear action potentials (CAP) and the 1-microV cochlear microphonic for 2-40 kHz tones, and the N1 amplitude intensity function (40-90 dB SPL). Cochlear histopathology was assessed in midmodiolar preparations of a separate set of animals, exposed as before (n = 4/group). RMA testing confirmed a TCE-induced loss in midfrequency threshold sensitivity (8 and 16 kHz). CAP thresholds were elevated at midfrequencies (8 and 16 kHz) among TCE-treated subjects, along with a suppression of the N1 amplitude from 50 to 90 dB SPL. The cochlear microphonic, a nonpropagated ac potential generated largely by the outer hair cells, was not affected by the TCE treatment. Cochlear histopathology revealed a loss of spiral ganglion cells that was significant in the middle turn, but not in the basal turn. There was an inconsistent loss of hair cells among treated subjects. The data suggest strongly that the behaviorally determined loss in auditory function can be accounted for by a cochlear impairment and that the spiral ganglion cell may be a prominent target of TCE.

Animals↗

PubMed1997

Cochlear protection from carbon monoxide exposure by free radical blockers in the guinea pig.

Acute carbon monoxide exposure produces a significant impairment in high-frequency auditory sensitivity that can be prevented using the N-methyl-D-aspartate receptor blocker MK-801. This finding suggests an excitotoxic component to carbon monoxide ototoxicity and establishes the potential for free radical formation. Free radical scavengers and inhibitors are protective in many organs, including the brain and cochlea, during hypoxic events such as ischemia/reperfusion and, in the cochlea, during noise exposure. This study evaluated the protection afforded by two such agents, phenyl-n-tert-butyl-nitrone (PBN), which acts as a general free radical scavenger, and allopurinol, which acts as a free radical inhibitor specific to the xanthine oxidase metabolic pathway. Guinea pigs were pretreated with PBN (100 mg/kg i.p.), allopurinol (100 mg/kg i.p.), or saline 1 hr prior to exposure to carbon monoxide (35 ml/kg i.p.) or to an equal volume of air. They were monitored at 15, 30, and 60 min after carbon monoxide exposure for alterations in compound action potential threshold and cochlear microphonic amplitude. The groups receiving carbon monoxide alone displayed characteristic compound action potential threshold elevations particularly at the higher test frequencies (16-40 kHz), consistent with earlier studies; no loss of cochlear microphonic amplitude was exhibited. Both free radical inhibitors, PBN and allopurinol, blocked loss of auditory threshold sensitivity produced by carbon monoxide. These data suggest that free radical generation may play a significant role in the impairment of high-frequency auditory sensitivity resulting from carbon monoxide.

Allopurinol↗

PubMed1997

Toluene disrupts outer hair cell morphometry and intracellular calcium homeostasis in cochlear cells of guinea pigs.

The aromatic hydrocarbon, toluene, has been demonstrated to disrupt auditory system function both in occupational epidemiological and in laboratory animal investigations. This agent, along with several other organic solvents, impairs hearing preferentially at middle frequencies-a finding that distinguishes these agents from the traditional high-frequency impairment observed with ototoxic drugs such as aminoglycoside antibiotics and cisplatin. Prior investigations have identified the outer hair cell as a probable target for toluene exposure, although studies designed to evaluate spiral ganglion cell impairment have not been reported. The purpose of this investigation was to determine directly whether outer hair cells isolated from the guinea pig cochlea show morphological alterations consistent with a toxic response to toluene exposure. Since slow adjustments of outer hair cell length can result from alteration in free intracellular calcium concentration, the effect of toluene on calcium homeostasis was monitored in both outer hair cells and spiral ganglion cells. A dose-response relationship was observed in the extent of outer hair cell shortening produced by toluene with a significant shortening observed at concentrations of 100 microM and higher. By contrast, the nonototoxic solvent, benzene, produced little shortening at 100 microM to 1 mM concentrations. Studies of calcium homeostasis conducted using the fluorescent probe, Fura-2, showed that toluene enhanced free intracellular calcium levels of both outer hair cells and spiral ganglion cells within 5 min of exposure at concentrations of 30 microM and higher. Intracellular calcium levels were elevated only slightly following benzene administration at 1 mM, but not at lower concentrations. Cells cultured in artificial perilymph nominally containing no calcium and those to which EGTA was added still showed a maximal increase in intracellular calcium level when treated with toluene. These data indicate that the elevation in free intracellular calcium levels produced by toluene results from release of calcium from intracellular stores.

Animals↗

PubMed1997

Correspondence between middle frequency auditory loss in vivo and outer hair cell shortening in vitro.

The aromatic hydrocarbon, toluene, has been reported to disrupt auditory system function both in occupational epidemiological and in laboratory animal investigations. This agent, along with several other organic solvents, impairs hearing preferentially at middle frequencies - a finding that distinguishes these agents from the traditional high frequency impairment observed with ototoxic drugs such as aminoglycoside antibiotics and cisplatin. Prior investigations performed in vivo have identified the outer hair cell as a probable target for toluene exposure. The purpose of this investigation was to determine directly whether outer hair cells isolated from the guinea pig cochlea show morphological alterations consistent with the toxic response seen in physiological studies with toluene exposure. The effect of toluene superfusion on outer hair cell shortening was assessed for cells harvested from different locations within the cochlea. Control studies included assessment of cell shortening among outer hair cells exposed to trimethyltin and cells exposed to benzene. Trimethyltin disrupts high frequency hearing preferentially and benzene does not produce hearing loss in vivo. Toluene at a concentration of 100 microM produced a marked shortening of outer hair cells although the effect was significantly greater among cells isolated from the apical half of the cochlea than from the basal half of the cochlea. By contrast, trimethyltin at the same concentration produced a preferential shortening among outer hair cells from the base of the cochlea. Benzene (100 microM) did not disrupt outer hair cell length of cells harvested from the apex. The results indicate that intrinsic features of outer hair cells contribute significantly to the site of ototoxic impairment observed in vivo for toluene.

Animals↗

PubMed1996

Comparison of the effects of trimethyltin on the intracellular calcium levels in spiral ganglion cells and outer hair cells.

Cochlear impairment by trimethyltin chloride (TMT), a potential contaminant of marine paints and polyvinyl chloride tubing, has been well demonstrated. Its toxic effect on the inner hair cells (IHC)-spiral ganglion cell (SGC) unit occurs almost immediately while disruption of outer hair cell (OHC) function does not occur until several hours after exposure. In this experiment, OHCs and SGCs from pigmented guinea pigs were tested in vitro to determine the role of enhanced intracellular calcium [Ca2+]i levels in TMT ototoxicity and to determine the sources of enhanced [Ca2+]i. The latter was determined by experiments using artificial perilymph without Ca2+ and by use of the Ca2+ channel blocker, nifedipine. The data show that TMT elevates [Ca2+]i in both OHC and SGC. The elevation of [Ca2+]i in SGC is much more rapid and larger than that in OHC. The elevation of [Ca2+]i in SGC can be attenuated by removing Ca2+ from artificial perilymph or pretreating with nifedipine, but neither of these treatments is effective in OHC. The results suggest that TMT disrupts intracellular storage of Ca2+ in OHCs and SGCs, but that is also enhancing influx of Ca2+ from extracellular sources in the SGCs.

Animals↗

PubMed1995

MK-801 protects against carbon monoxide-induced hearing loss.

MK-801, a glutamate receptor blocker, has been reported to protect against hypoxic-ischemic neuronal degeneration. The purpose of this study was to investigate the protective effects of MK-801 on cochlear impairment induced by carbon monoxide (CO) in the guinea pig and whether the protection reflected systemic or local effects. Glutamate has been proposed to be the neurotransmitter at the inner hair cell/type I spiral ganglion cell synapse. CO (35 ml/kg ip) elevated the compound action potential threshold at high frequencies (16-40 kHz) 30 min after treatment. In the group pretreated with 1 mg/kg MK-801 ip there was no significant elevation of compound action potential threshold 30 min after CO exposure, suggesting that MK-801 provided some protection at this dosage. In the group pretreated with 0.1 mg/kg MK-801 there was protection against cochlear dysfunction 15 min after CO injection, but significant elevation of compound action potential threshold occurred 30 and 60 min after CO exposure. Data also indicated that 1 mM MK-801 applied topically on the round window membrane provided protective effects against CO hypoxia from 15 up to 60 min. This experiment suggests that cochlear impairment induced by CO hypoxia may result from excess extracellular concentrations of glutamate.

Animals↗

PubMed1995

Elevation of intracellular calcium levels in spiral ganglion cells by trimethyltin.

The neurotoxicant, trimethyltin (TMT) produces cochlear impairment at far lower dose levels and far more rapidly than it does central nervous system effects. The initial effects of TMT in the cochlea, in vivo, are consistent with disruption of the inner hair cell type-1 spiral ganglion cell synapse although it is uncertain whether the effect is on presynaptic and/or postsynaptic units. This synapse is believed to be an excitatory glutamatergic one, providing the possibility that TMT could induce an excitotoxic process resulting in elevations in intracellular calcium ([Ca2+]i). The objective of this study was to determine whether TMT had direct toxic effects on the postsynaptic spiral ganglion cells studied in primary culture and to identify the role of extracellular calcium in such an effect. The marker of interest was the effect of this agent on [Ca2+]i levels as determined using quantitation of the fluorescent calcium dye, Fura-2. TMT did induce a marked and sustained elevation in [Ca2+]i level in the spiral ganglion cells that appeared to have a rapid initial phase and a slower saturating phase. Studies performed using calcium-free medium showed that elevation of [Ca2+]i in spiral ganglion cells by TMT was attenuated but not entirely blocked. Further, the L-type calcium channel blocker, nifedipine, was able to inhibit the initial increase in [Ca2+]i, suggesting that at least this phase of the TMT effect was mediated by calcium channels, although nifedipine had no significant effect on the time to reach the maximal [Ca2+]i level. Parallel control experiments performed using application of exogenous glutamate and depolarizing K+ concentrations also produced elevation in [Ca2+]i levels. The data indicate that TMT elevates [Ca2+]i in isolated spiral ganglion cells both by increasing extracellular uptake via Ca2+ channels and also by releasing Ca2+ from intracellular stores. Thus TMT ototoxicity appears to include a direct postsynaptic toxic event.

Analysis of Variance↗