Toluene-induced hearing loss in phenobarbital treated rats.

Exposure to aromatic organic solvents may induce hearing loss in rats, the cochlea being the primary target. The aim of this study which was carried out in rat, was to evaluate the impact of the hepatic metabolism of toluene on its ototoxic potency. To this end, the solvent hepatic metabolism was shifted by treating the rats with 50 mg/kg/d of phenobarbital (PhB), a potent inducer of the microsomal cytochromes P450 system, alcohol and aldehyde dehydrogenases, and glutathione-S-transferases. The two main urinary metabolites of the oxidative and conjugate pathways [hippuric (HA) and benzyl mercapturic acids (BMA) respectively] confirmed the efficacy of the PhB treatment. For the PhB-induced rats, the amount of excreted HA increased by 43% and the amount of BMA by 35%. Auditory function impairments were assessed using auditory-evoked potentials. On completion of the auditory tests, the organs of Corti were dissected to evaluate hair cell losses. The permanent auditory threshold shifts were approximately 15 dB greater in the toluene-exposed rats than in the PhB-induced rats. Both the functional and morphological data confirmed that PhB treatment can decrease the ototoxic potency of toluene.

Effects of aromatic solvents on acoustic reflexes mediated by central auditory pathways.

From previous in vivo investigations, it has been shown that toluene can mimic the effects of cholinergic receptor antagonists and may thereby modify the response of protective acoustic reflexes. The current study aimed to define the relative effects of aromatic solvents on the middle ear and inner ear acoustic reflexes. Toward this end, the cochlear microphonic (CMP) elicited with a band noise centered at 4 kHz, and the compound action potential (CAP) elicited with 4-kHz tone pips was measured in rats. Both potentials were recorded before, during, and after triggering the protective reflexes by a 110-dB SPL contralateral octave band noise centered at 12.5 kHz (12.5 kHz-OBN). In several rats, the middle ear muscles were severed to identify the relative effects of toluene on the two reflexes. While the reflex elicitor was capable of decreasing both the CMP and CAP amplitudes, an injection of 116.2 mM toluene cancelled this suppressor effect induced by the contralateral sound. In the rats with nonfunctional middle ear muscles, a solvent injection did not modify the electrophysiological responses of the cochlea. Different solvents were tested to study the relationship of the chemical structure of the solvents on the acoustic reflexes. The present study showed that aromatic solvents can inhibit the action of the middle ear reflex by their anticholinergic effect on the efferent motoneurons. An aromatic nucleus and the presence of one side chain of no more than 3 C seem to be required in the solvent structure to inhibit the efferent motoneurons.

Increase in cochlear microphonic potential after toluene administration.

Human and animal studies have shown that toluene can cause hearing loss. In the rat, the outer hair cells are first disrupted by the ototoxicant. Because of their particular sensitivity to toluene, the cochlear microphonic potential (CMP) was used for monitoring the cochlea activity of anesthetized rats exposed to both noise (band noise centered at 4 kHz) and toluene. In the present experiment, the conditions were specifically designed to study the toluene effects on CMP and not those of its metabolites. To this end, 100-microL injections of a vehicle containing different concentrations of solvent were made into the carotid artery connected to the tested cochlea. Interestingly, an injection of 116.2-mM toluene dramatically increased in the CMP amplitude (approximately 4 dB) in response to an 85-dB SPL noise. Moreover, the rise in CMP magnitude was intensity dependent at this concentration suggesting that toluene could inhibit the auditory efferent system involved in the inner-ear or/and middle-ear acoustic reflexes. Because acetylcholine is the neurotransmitter mediated by the auditory efferent bundles, injections of antagonists of cholinergic receptors (AchRs) such as atropine, 4-diphenylacetoxy-N-methylpiperidine-methiodide (mAchR antagonist) and dihydro-beta-erythroidine (nAchR antagonist) were also tested in this investigation. They all provoked rises in CMP having amplitudes as large as those obtained with toluene. The results showed for the first time in an in vivo study that toluene mimics the effects of AchR antagonists. It is likely that toluene might modify the response of protective acoustic reflexes.

Ototoxicity of the three xylene isomers in the rat.

Numerous experiments have shown that the aromatic solvents can affect the auditory system in the rat, the cochlea being targeted first. Solvents differ in cochleotoxic potency: for example, styrene is more ototoxic than toluene or xylenes. The goal of this study was to determine the relative ototoxicity of the three isomers of xylene (o-, m- or p-xylene). Moreover, by dosing with the two urinary metabolites of xylene, methylhippuric (MHAs) and mercapturic acids (MBAs), this study points toward a causal relationship between the cochleotoxic effects and potential reactive intermediates arising from the biotransformation of the parent molecules. Separate groups of rats were exposed by inhalation to one isomer following this schedule: 1800 ppm, 6 h/d, 5 d/wk for 3 wk. Auditory thresholds were determined with brainstem-auditory evoked potentials. Morphological analysis of the organ of Corti was performed by counting both sensory and spiral ganglion cells. Among the three isomers, only p-xylene was cochleotoxic. A 39-dB permanent threshold shift was obtained over the tested frequencies range from 8 to 20 kHz. Whereas outer hair cells were largely injured, no significant morphological change was observed within spiral ganglia. The concentrations of urinary p-, o- or m-MHA were greater (p-MHA: 33.2 g/g; o-MHA: 7.8 g/g; m-MHA: 20.4 g/g) than those obtained for MBAs (p-MBA: 0.04 g/g; o-MBA: 6.2 g/g; m-MBA: 0.03 g/g). Besides, there is a large difference between o-MBA (6.2 g/g) and p-MBA (0.04 g/g). As a result, since the cysteine conjugates are not determinant in the ototoxic process of xylenes, the location of the methyl groups around the benzene nucleus could play a key role.

Influence of age on noise- and styrene-induced hearing loss in the Long-Evans rat.

This paper reviews different investigations carried out with Long-Evans rats on the influence of age on the ototoxicity induced by styrene and on the vulnerability to noise. The first part of this article is focused on the differences in auditory susceptibility to noise (92 or 97dB octave band noise centered at 8kHz, 6 h/day, 5 days/week, 4 weeks) and styrene (700ppm, 6h/d, 5 d/w, 4 w) between young (three and half months) and old (24 months) Long-Evans rats. Auditory evoked potential measures revealed that the old rats tend to be more sensitive than young rats to higher noise levels (97dB), but equally vulnerable to moderate levels (92dB). By contrast, the aged rats were virtually insensitive to 700ppm styrene compared to the young animals. Two additional studies were performed controlling and examining the influence of body weight and post-natal age on the sensitivity to styrene. Rats of the same age (21 weeks) and but having different body weight (∼310g versus ∼410g) did not show any difference of sensitivity to 700ppm styrene, whereas 14-week-old rats with the same body weight as 21-week-old rats (∼350g) revealed increased sensitivity to styrene. These results show that weight does not play a key role in the sensitivity to styrene, and suggest a long period of increased sensitivity to styrene during the first months of life.

Solvent ototoxicity in the rat and guinea pig.

There is clear evidence that aromatic solvents can disrupt the auditory system in humans and animals. As far as animal models are concerned, solvent-induced hearing loss seems to be species-dependent. Indeed, most published data have been obtained with the rat, which shows mid-frequency cochlear deficits, whereas the guinea pig does not show any permanent hearing loss after solvent exposure. In the current investigation, the effects of two solvents, toluene (600 ppm) and styrene (1000 ppm), were studied in both Long-Evans rats and pigmented guinea pigs exposed 6 h/day for 5 consecutive days. Cochlear function was tested by using distortion product otoacoustic emissions (DPOAE) measured prior to the solvent exposure, 20 min after the end of the exposure and successively at 2 and 4 weeks post-exposure. In addition to cochlear testing, solvent concentrations in blood and urinary metabolites were measured. A cochlear histological analysis was performed at the end of the experiment. No decrease in DPOAE amplitude was observed in the guinea pig, even immediately following the end of exposure. The rat model showed severe disruption of auditory function and cochlear pathology, whereas the guinea pig had no disruption of DPOAE or cochlear pathological alterations. Therefore, the vulnerability of the cochlear function was strictly dependent on the species. As expected, an important difference in the styrene concentration in blood was observed: the solvent concentrations were fourfold higher in the rat than in the guinea pig. Therefore, it is clear that a pharmacokinetic or an uptake difference might explain the difference in susceptibility observed between the two species. Moreover, the metabolism pathways of the solvents were different depending on the species. Attempts to explain differences of vulnerability between the rat and guinea pig are addressed in the present paper.

Use of DPOAEs for assessing hearing loss caused by styrene in the rat.

The study was carried out to test whether or not cubic distortion otoacoustic emissions were more sensitive than auditory-evoked potentials for assessing styrene-induced hearing losses in the Long-Evans rat. For the purposes of comparison, changes in cubic distortion product otoacoustic emissions (DeltaDPOAE), evoked potential permanent threshold shifts (PTS) and outer hair cell losses were measured in a population of styrene-treated rats. Each rat was exposed to either 650 or 750 ppm of styrene for 4 weeks, 5 days per week, 6 h per day. Only the 750 ppm exposure caused significant hearing losses. For this concentration, DPOAEs appeared as sensitive to styrene as the audiometry performed with evoked potentials, but not more. A high coefficient of correlation [0.84< or =r< or =0.91] between DeltaDPOAE and PTS was obtained across the styrene-induced effects for frequencies ranging from 5 to 12 kHz. This experiment demonstrates that DPOAEs can be used to monitor the ototoxicity induced by styrene even though they cannot be considered as a more sensitive index of cochlear pathology than the evoked potentials, at least under our experimental conditions. Likewise evoked potentials, normal DPOAEs may not guarantee a normal cochlear status and therefore results of DPOAE measurements should be interpreted cautiously. The use of both techniques and the determination of the ratio DeltaDPOAE/PTS may be useful in determining the cause of hearing loss: mechanical or chemical process. Moreover, because of its non-invasive and objective characteristics, the use of DPOAEs could play a greater role in a prevention policy.