Toluene can perturb the neuronal voltage-dependent Ca2+ channels involved in the middle-ear reflex.

Numerous laboratory-based data have shown the ability of toluene (Tol) to exacerbate noise-induced hearing loss. However, the mechanism responsible for the synergistic effects of a coexposure to noise and Tol has not yet been completely elucidated. Recent investigations in rats have focused on quantifying the anticholinergic effects of certain aromatic solvents and have demonstrated that these solvents can cancel the protective role played by the middle-ear reflex (MER). Voltage-dependent Ca(2+) channels (VDCCs) regulate acetylcholine release in the central synaptic network and control muscular excitation/contraction processes as well. In order to identify the prevailing action of Tol in the central or peripheral compartment of the MER arc, two VDCC antagonists were injected into the common carotid trunk: omega-conotoxin MVIIC, which blocks only the neuronal N- and P/Q-type Ca(2+) channels, or verapamil, which inhibits the muscular L-type Ca(2+) channels. Rats were also implanted with an electrode on the round window membrane to measure the cochlear microphonic potential (CMP) elicited with a band noise centered at 4 kHz and emitted at 85 dB sound pressure level. The variations in CMP recorded during the test compound injection showed that Tol has similar effects to those induced by omega-conotoxin, the neuronal VDCC blocker. The response obtained with the verapamil injection was broader than those obtained with Tol or conotoxin. This investigation therefore revealed that Tol can mimic the effects of VDCC blockers. The antagonist effects of Tol would be closer to neuronal than to muscular blockers and would be presumably located at the level of the integrator centers of the reflex.

Solvent-induced hearing loss: mechanisms and prevention strategy.

While noise exposure is the most significant contributor to occupational hearing loss, evidence gained over the last 10 years, has pointed to organic solvents as additional contributors to occupational hearing disorders. Despite the implications of this finding, no significant measure has been undertaken to limit exposure to occupational solvents, or to occupational solvents and noise, within the European community. Guidelines for improving hearing protection of people exposed to solvents, or to solvents and noise, are addressed in the present article. Recently, it has been shown that the lowest-observed-adverse-effect level (LOAEL) of styrene was 300 ppm in active (working wheel) rats, and that the same amount of styrene-induced hearing loss (SIHL) can be obtained with styrene concentration difference of 200 ppm between active and sedentary (inactive) rats. Supported by a reasonable safety factor (SF) of 10, the authors proposed to decrease the French threshold limit value of styrene from 50 to 30 ppm (RfD=LOAEL/SF) to ensure a higher level of protection for human hearing. It is widely acknowledged that outer hair cells in the organ of Corti can be considered as the first target tissue of solvents, while little is known about the action of aromatic solvents on the auditory efferent system. In a recent experiment using both the cochlear microphonic and compound action potentials, the authors have shown that toluene can inhibit the action of the middle ear reflex by modifying the cholinergic receptors. It is likely that toluene affects the cholinergic receptors at the brainstem level. By its anticholinergic-like effect, toluene could allow higher acoustic energy penetration into the cochlea exposed to both noise and solvent. Based on this phenomenon, the authors recommend the use of hearing protection for the lower exposure action value: Lex,8h=80 dB(A) in noisy environments polluted by solvents.

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.

Consequences of noise- or styrene-induced cochlear damages on glutamate decarboxylase levels in the rat inferior colliculus.

Both noise and styrene can injure the cochlea, resulting in a reduction of incoming inputs from the cochlea to the central nervous system. In addition, styrene is known to have neurotoxic properties at high doses. The loss of inputs caused by noise has been shown to be compensated by a new equilibrium between excitatory and inhibitory influences within the inferior colliculus (IC). The main goal of this study was to determine whether styrene-induced hearing loss could also be counterbalanced by a GABAergic adjustment in the IC. For this purpose, rats were exposed to noise (97 dB SPL octave band noise centered at 8 kHz), or to a non-neurotoxic dose of styrene for 4 weeks (700 ppm, 6 h/day, 5 days/week). Auditory sensitivity was tested by evoked potentials, and cochlear damage was assessed by hair cell counts. Glutamate decarboxylase (GAD) was dosed in the IC by indirect competitive enzyme-linked immunosorbent assay. Both noise and styrene caused PTSs that reached 27.0 and 14.6 dB respectively. Outer hair cell (OHC) loss caused by noise did not exceed 9% in the first row, on the other hand OHC loss induced by styrene reached 63% in the third row. Only the noise caused a decrease of GAD of 37% compared to that measured in the controls. No significant modification of GAD concentration has been shown after styrene exposure. Thus, central compensation for cochlear damage may depend on the nature of the ototoxic agent. Unless styrene directly affects IC function, it is reasonable to assume that noise causes a modification of inhibitory neurotransmission within the structure because of impairment of afferent supply to the auditory brainstem. The present findings suggest that central compensation for cochlear damage can preferably occur when afferent fibers are altered.