High-intensity Sound Stimulation Research Articles

Cannabinoids in Audiogenic Seizures: From Neuronal Networks to Future Perspectives for Epilepsy Treatment.

Cannabinoids and Cannabis-derived compounds have been receiving especial attention in the epilepsy research scenario. Pharmacological modulation of endocannabinoid system's components, like cannabinoid type 1 receptors (CB1R) and their bindings, are associated with seizures in preclinical models. CB1R expression and functionality were altered in humans and preclinical models of seizures. Additionally, Cannabis-derived compounds, like cannabidiol (CBD), present anticonvulsant activity in humans and in a great variety of animal models. Audiogenic seizures (AS) are induced in genetically susceptible animals by high-intensity sound stimulation. Audiogenic strains, like the Genetically Epilepsy Prone Rats, Wistar Audiogenic Rats, and Krushinsky-Molodkina, are useful tools to study epilepsy. In audiogenic susceptible animals, acute acoustic stimulation induces brainstem-dependent wild running and tonic-clonic seizures. However, during the chronic protocol of AS, the audiogenic kindling (AuK), limbic and cortical structures are recruited, and the initially brainstem-dependent seizures give rise to limbic seizures. The present study reviewed the effects of pharmacological modulation of the endocannabinoid system in audiogenic seizure susceptibility and expression. The effects of Cannabis-derived compounds in audiogenic seizures were also reviewed, with especial attention to CBD. CB1R activation, as well Cannabis-derived compounds, induced anticonvulsant effects against audiogenic seizures, but the effects of cannabinoids modulation and Cannabis-derived compounds still need to be verified in chronic audiogenic seizures. The effects of cannabinoids and Cannabis-derived compounds should be further investigated not only in audiogenic seizures, but also in epilepsy related comorbidities present in audiogenic strains, like anxiety, and depression.

Loss of Brain-Derived Neurotrophic Factor Mediates Inhibition of Hippocampal Long-Term Potentiation by High-Intensity Sound.

Exposure to noise produces cognitive and emotional disorders, and recent studies have shown that auditory stimulation or deprivation affects hippocampal function. Previously, we showed that exposure to high-intensity sound (110dB, 1min) strongly inhibits Schaffer-CA1 long-term potentiation (LTP). Here we investigated possible mechanisms involved in this effect. We found that exposure to 110dB sound activates c-fos expression in hippocampal CA1 and CA3 neurons. Although sound stimulation did not affect glutamatergic or GABAergic neurotransmission in CA1, it did depress the level of brain-derived neurotrophic factor (BDNF), which is involved in promoting hippocampal synaptic plasticity. Moreover, perfusion of slices with BDNF rescued LTP in animals exposed to sound stimulation, whereas BDNF did not affect LTP in sham-stimulated rats. Furthermore, LM22A4, a TrkB receptor agonist, also rescued LTP from sound-stimulated animals. Our results indicate that depression of hippocampal BDNF mediates the inhibition of LTP produced by high-intensity sound stimulation.

Long-term high-intensity sound stimulation inhibits h current (Ih ) in CA1 pyramidal neurons.

Afferent neurotransmission to hippocampal pyramidal cells can lead to long-term changes to their intrinsic membrane properties and affect many ion currents. One of the most plastic neuronal currents is the hyperpolarization-activated cationic current (Ih ), which changes in CA1 pyramidal cells in response to many types of physiological and pathological processes, including auditory stimulation. Recently, we demonstrated that long-term potentiation (LTP) in rat hippocampal Schaffer-CA1 synapses is depressed by high-intensity sound stimulation. Here, we investigated whether a long-term high-intensity sound stimulation could affect intrinsic membrane properties of rat CA1 pyramidal neurons. Our results showed that Ih is depressed by long-term high-intensity sound exposure (1min of 110 dB sound, applied two times per day for 10days). This resulted in a decreased resting membrane potential, increased membrane input resistance and time constant, and decreased action potential threshold. In addition, CA1 pyramidal neurons from sound-exposed animals fired more action potentials than neurons from control animals; however, this effect was not caused by a decreased Ih . On the other hand, a single episode (1min) of 110 dB sound stimulation which also inhibits hippocampal LTP did not affect Ih and firing in pyramidal neurons, suggesting that effects on Ih are long-term responses to high-intensity sound exposure. Our results show that prolonged exposure to high-intensity sound affects intrinsic membrane properties of hippocampal pyramidal neurons, mainly by decreasing the amplitude of Ih .

Oxidative stress and Na,K-ATPase activity differential regulation in brainstem and forebrain of Wistar Audiogenic rats may lead to increased seizure susceptibility

The Wistar Audiogenic Rat (WAR) is a well-characterized seizure-prone, inbred rodent strain that, when acutely stimulated with high-intensity sounds, develops brainstem-dependent tonic-clonic seizures that can evolve to limbic-like, myoclonic (forebrain) seizures when the acoustic stimuli are presented chronically (audiogenic kindling). In order to investigate possible mechanisms underlying WAR susceptibility to seizures, we evaluated Na,K-ATPase activity, Ca-ATPase activity, Mg-ATPase activity, lipid membrane composition and oxidative stress markers in whole forebrain and whole brainstem samples of naïve WAR, as compared to samples from control Wistar rats. We also evaluated the expression levels of α1 and α3 isoforms of Na,K-ATPase in forebrain samples. We observed increased Na,K-ATPase activity in forebrain samples and increased oxidative stress markers (lipid peroxidation, glutathione peroxidase and superoxide dismutase) in brainstem samples of WAR. The Ca-ATPase activity, Mg-ATPase activity, lipid membrane composition and expression levels of α1 and α3 isoforms of Na,K-ATPase were unaltered. In view of previous data showing that the membrane potentials from naïve WAR’s neurons are less negative than that from neurons from Wistar rats, we suggest that Na,K-ATPase increased activity might be involved in a compensatory mechanism necessary to maintain WAR’s brains normal activity. Additionally, ongoing oxidative stress in the brainstem could bring Na,K-ATPase activity back to normal levels, which may explain why WAR’s present increased susceptibility to seizures triggered by high-intensity sound stimulation.

Wistar audiogenic rats display abnormal behavioral traits associated with artificial selection for seizure susceptibility.

Accumulating evidence from different animal models has contributed to the understanding of the bidirectional comorbidity associations between the epileptic condition and behavioral abnormalities. A strain of animals inbred to enhance seizure predisposition to high-intensity sound stimulation, the Wistar audiogenic rat (WAR), underwent several behavioral tests: forced swim test (FST), open-field test (OFT), sucrose preference test (SPT), elevated plus maze (EPM), social preference (SP), marble burying test (MBT), inhibitory avoidance (IAT), and two-way active avoidance (TWAA). The choice of tests aimed to investigate the correlation between underlying circuits believed to be participating in both WAR's innate susceptibility to sound-triggered seizures and the neurobiological substrates associated with test performance. Comparing WAR with its Wistar counterpart (i.e., resistant to audiogenic seizures) showed that WARs present behavioral despair traits (e.g., increased FST immobility) but no evidence of anhedonic behavior (e.g., increased sucrose consumption in SPT) or social impairment (e.g., no difference regarding juvenile exploration in SP). In addition, tests suggested that WARs are unable to properly evaluate degrees of aversiveness (e.g., performance on OFT, EPM, MBT, IAT, and TWAA). The particularities of the WAR model opens new venues to further untangle the neurobiology underlying the co-morbidity of behavioral disorders and epilepsy. This article is part of a Special Issue entitled "Genetic and Reflex Epilepsies, Audiogenic Seizures and Strains: From Experimental Models to the Clinic".

Inhibition of long-term potentiation in the schaffer-CA1 pathway by repetitive high-intensity sound stimulation

High-intensity sound can induce seizures in susceptible animals. After repeated acoustic stimuli changes in behavioural seizure repertoire and epileptic EEG activity might be seen in recruited limbic and forebrain structures, a phenomenon known as audiogenic kindling. It is postulated that audiogenic kindling can produce synaptic plasticity events leading to the spread of epileptogenic activity to the limbic system. In order to test this hypothesis, we investigated if long-term potentiation (LTP) of hippocampal Schaffer-CA1 synapses and spatial navigation memory are altered by a repeated high-intensity sound stimulation (HISS) protocol, consisting of one-minute 120dB broadband noise applied twice a day for 10days, in normal Wistar rats and in audiogenic seizure-prone rats (Wistar Audiogenic Rats – WARs). After HISS all WARs exhibited midbrain seizures and 50% of these animals developed limbic recruitment, while only 26% of Wistar rats presented midbrain seizures and none of them had limbic recruitment. In naïve animals, LTP in hippocampal CA1 neurons was induced by 50- or 100-Hz high-frequency stimulation of Schaffer fibres in slices from both Wistar and WAR animals similarly. Surprisingly, HISS suppressed LTP in CA1 neurons in slices from Wistar rats that did not present any seizure, and inhibited LTP in slices from Wistar rats with only midbrain seizures. However HISS had no effect on LTP in CA1 neurons from slices of WARs. Interestingly HISS did not alter spatial navigation and memory in both strains. These findings show that repeated high-intensity sound stimulation prevent LTP of Schaffer-CA1 synapses from Wistar rats, without affecting spatial memory. This effect was not seen in hippocampi from audiogenic seizure-prone WARs. In WARs the link between auditory stimulation and hippocampal LTP seems to be disrupted which could be relevant for the susceptibility to seizures in this strain.

Potencial evocado miogênico vestibular (Vemp): avaliação das respostas em indivíduos normais

the Vestibular Evoked Myogenic Potential (Vemp) is formed by myogenic neurophysiologic responses activated by high-intensity sound stimulation. The response is registered through surface electromyography of the cervical muscles during muscle contraction. The acoustic stimuli activate the saccular macula, the vestibular inferior nerve and the pathways related to the vestibule-spinal descendant nerves. to describe Vemp parameters in a normal population. thirty adults, 13 men and 17 women with no otoneurological complaints were selected. The stimuli were 200 tone burst, with a frequency of 1Hz and intensity of 118 dB Na, band-pass filter ranging from 10Hz to 1500Hz. The first potential biphasic P13-N23 wave was analyzed. no significant difference was observed between the sides of stimulation in terms of latency and amplitude. However, a statistically significant difference was found for amplitude between genders. Vemp demonstrated to be a reliable instrument in the clinical assessment of the vestibular function.

Age-Related Changes in Vestibular Evoked Myogenic Potentials

Vestibular evoked myogenic potentials (VEMP) in response to sound stimulation (500 Hz tone burst, 129 dB SPL) were studied in 1000 consecutive patients. VEMP from the ear with the larger amplitude were evaluated based on the assumption that the majority of the tested patients probably had normal vestibular function in that ear. Patients with known bilateral conductive hearing loss, with known bilateral vestibular disease and those with Tullio phenomenon were not included in the evaluation. It was found that there was an age-related decrease in VEMP amplitude and an increase in VEMP latency that appeared to be rather constant throughout the whole age span. The VEMP data were also compared to an additional group of 10 patients with Tullio phenomenon. Although these 10 patients did have rather large VEMP, equally large VEMP amplitudes were observed in a proportion of unaffected subjects of a similar age group. Thus, the finding of a large VEMP amplitude in response to a high-intensity sound stimulation is not, per se, distinctive for a significant vestibular hypersensitivity to sounds.

Vasopressinergic hypothalamic neurons are recruited during the audiogenic seizure of WARs

The Wistar Audiogenic Rat (WAR) is a genetic model of reflex epilepsy with seizures induced by high-intensity sound stimulation (120 dB SPL). In spite of the known neural substrates involved in WAR seizure phenotype, neuroendocrine hypothalamic neurons were never investigated. In this work, AVP immunohistochemistry in the hypothalamus and radioimmunoassay (RIA) in plasma and in hypothalamic and hypophysial tissues were performed on both controls and WARs in order to evaluate the dynamics of AVP release due to seizure induction. Susceptible animals (WARs) displayed at least tonic–clonic convulsions followed by clonic spasms, while resistant Wistar rats (R) had no convulsive behavior. Animals were sacrificed at 3 instances: basal condition (without stimulus) and at 3 and 10 min after sound stimulation. For the immunohistochemistry AVP study, brains were harvested and processed by the avidin–biotin–peroxidase detection method. Optic densitometry was used for quantifying AVP labeling in supraoptic (SON) and paraventricular (PVN) hypothalamic nuclei. SON presented higher densitometry levels (% D—relative to background) for both WARs and R when compared to PVN. Nevertheless, both nuclei presented a marked decrease, referenced to basal levels, in % D for WARs at 3 min (~35%) against a discrete change for R (~90%). RIA results were significantly higher in the hypophysis of WARs when compared to R rats, at 3 min. Also, at 3 min, plasma AVP in WARs (89.32 ± 24.81 pg/mL) were higher than in R (12.01 ± 2.39 pg/mL). We conclude, based on the AVP releasing profiles, that vasopressinergic hypothalamic neurons are recruited during the audiogenic seizure of WARs.

Evidence for augmented brainstem activated forebrain seizures in Wistar Audiogenic Rats subjected to transauricular electroshock

Previous work from our laboratory has shown that naïve Wistar Audiogenic Rats (WARs), a genetic model of reflex epilepsy in which seizures are induced by high-intensity sound stimulation (120 dB SPL), are seizure-prone to a variety of pro-convulsive stimuli (e.g., transauricular electroshock, pentylenetetrazole and pilocarpine). On the other hand, repetitive acoustic stimulation of WARs causes a slow recruitment of limbic structures, known as audiogenic kindling, changing seizure expression to include behavior characteristic of temporal-lobe epilepsy. Thus, our hypothesis is that WARs have facilitated acoustic-limbic projections when compared to Wistar controls. Wistar controls ( n = 9) and WARs ( n = 9) underwent EEG electrode implants in the cortex-Cx, amygdaloid complex-AMY and inferior colliculus-IC and received one low current transauricular electrical stimulus (ES) daily, for three consecutive days, with intensities of 10, 20 and 30 mA, respectively. The video-electroencephalographic activity was recorded 1 min before and 4 min after ES. Our results confirm previously described data indicating a greater susceptibility of WARs to seizure. However, low current ES (e.g., 20 mA) triggered epileptiform activity in the AMY only after epileptiform EEG was visible in the Cx and IC electrode leads. The AMY after-discharge continued even though no evident epileptiform activity was present in the Cx. In conclusion, our results add electrophysiological data to previously published behavioral evidence of WAR enhanced susceptibility to ES seizures and, also, support the hypothesis that the acoustic-limbic circuitry is facilitated even in unkindled WARs.

Assessment of the seizure susceptibility of Wistar Audiogenic rat to electroshock, pentyleneterazole and pilocarpine

This work evaluates the seizure susceptibility of naı̈ve female Wistar Audiogenic rats (WARs), a genetic model of reflex epilepsy in which seizures are induced by high-intensity sound stimulation (120 dB SPL), to other pro-convulsive stimuli: transauricular electroshock (ES), pentylenetetrazole (PTZ) and pilocarpine (PILO). Normal Wistar rats from the main breeding stock of the Institute of Biological Sciences, UFMG were taken as controls. Electroshock seizures were induced through a pair of ear-clip electrodes (10 mA, at a frequency of 60 Hz, applied for 1 s). In order to test WAR susceptibility to chemically induced seizures, animals were treated either with PTZ (37.5 mg/kg i.p.) or PILO (200, 270 and 300 mg/kg i.p.). Seizure severity was evaluated by appropriate behavioral severity index scales (SI) specific to each epilepsy model and tested for statistical significance using the non-parametric Mann–Whitney Rank Sum test. Results show a significantly greater susceptibility of WARs for ES (SI WAR=3±3/3, SI Wistar=1±1/1; median±interquartile range 25%/75%, P<0.01) and PTZ (SI WAR=4±4/4, SI Wistar=1±1/4; median±interquartile range 25%/75%, P<0.02), as indicated by significantly higher SI scores and shorter latencies for seizure onset ( T WAR=71±7 s, T Wistar=94±8 s; P<0.05 Student t-test, mean±S.E.M.). Although PILO also caused higher SI scores in WARs (WAR 200 mg=1±1/1, Wistar 200 mg=1±1/1; WAR 270 mg=1.5±1/2, Wistar 270 mg=1±1/1.25; WAR 300 mg=9±1/9, Wistar 300 mg=4±1.5/7.5; median±interquartile range 25%/75%), statistically significant differences were not observed. In conclusion, our results show that WARs have an inherited broader predisposition for seizures.

Audiogenic kindling in the Wistar rat: a potential model for recruitment of limbic structures

Repetitive high intensity (110 dB) sound stimulation induces a forebrain-kindling phenomenon in animals predisposed to sound induced seizures. Wistar audiogenic rats (WARs) have been reported to develop a mixed brainstem-limbic seizure pattern, after more than five to ten stimuli. Besides the original brainstem wild running and tonic-clonic seizures, new behavioral patterns appear resembling those of electrical amygdala kindling. Although audiogenic kindling is a well-known phenomenon, electrographic limbic recruitment during the kindling has never been reported. Our objective was to use electrophysiology to test the hypothesis of gradual and sequential involvement of the amygdala and then cortex during audiogenic kindling. We used video-EEG recordings with cortical and deep electrode implants (inferior colliculus and basolateral amygdaloid nuclei) during audiogenic kindling on eight WARs, and their respective controls, submitted to a protocol of three acoustic stimuli per day. A new design for ‘on site’ source follower circuits was used in order to minimize noise during the recording of EEG data from the wild running episode and the subsequent tonic-clonic or motor limbic seizures. The video-EEG equipment assembled allowed synchronous recordings of both behavior and EEG. WARs first recordings showed electrodecremental responses after seizure onset and a probable epileptiform activity, particularly in the inferior colliculus, during the tonic phase of seizure. All animals showed very similar polyspike-wave activity in the amygdala, after behavioral seizure patterns (Racine’s scale) occurred. The morphology of such epileptiform EEG activity is very similar to that reported for electrical amygdala kindling. Also, when audiogenic kindling continued, both inferior colliculus and cortical electrodes presented high amplitude and synchronized epileptiform polyspike activity.

Genetic Mapping of a Locus (mass1) Causing Audiogenic Seizures in Mice

Frings audiogenic seizure-susceptible mice are a model for sensory-evoked reflex seizures. Their seizure phenotype is characterized by wild running, loss of righting reflex, tonic flexion, and tonic extension in response to high-intensity sound stimulation. The Frings mice represent an inbred colony that has not been genetically characterized. This investigation studied the mode of inheritance for audiogenic seizures by crossing the Frings mouse with the seizure-resistant C57BL/6J mouse. Among the backcross progeny generated by crossing (Frings × C57BL/6J)F1 mice with the Frings strain, 391 of the 836 N2 progeny were audiogenic seizure susceptible, a finding consistent with monogenic inheritance. Genetic mapping and linkage analysis of hybrid mice using MIT microsatellite marker sequences localized the seizure gene, namedmass1for monogenic audiogenic seizure susceptible, to an approximately 3.6 cM interval in the middle of mouse chromosome 13. Linkage ofmass1to chromosome 13 is an important step in identifying the gene associated with a monogenic seizure disorder in mice, which may ultimately lead to a better understanding of the pathophysiology of human seizure disorders.

An automated system for mapping spatial movement patterns in behavioral research

A computerized positioning system was developed and used to map gross body movement of children around a room in response to high-intensity sound stimulation. Details of the data collection system are discussed. The system has potential for use in a variety of research projects when it is necessary to identify position coordinates and analyze changes in spatial location for subjects within a test room environment.

NMDA-dependent audiogenic seizures are differentially regulated by inferior colliculus subnuclei

Wistar rats were classified as susceptible (S) and resistant (R) to audiogenic seizures (AS) by evaluation of their response to high intensity sound stimulation (110.3 dB). R rats usually do not respond with any convulsive behavior to sound stimulation, whereas S animals develop a complex wild running sequence plus tonic-clonic seizure patterns after sound stimulation. Thus, R rats were injected with phosphate buffer (PB; 0.2 μl) or N- methyl- d-aspartate (NMDA) in three different doses (2.0 μg, 2.5 μg and 3.0 μg/0.2 μl) into central ventral or cortical dorsal inferior colliculus (IC) nuclei. Dose-response curves were evaluated by means of an ethological method in which behavioral sequences typical of S and R animals were quantitated. Animals displayed more severe spontaneous audiogenic-like seizures with the dose of 2.5 μg/0.2 μl NMDA, which were potentiated by the acoustic stimulus. Significant differences were apparent between central and cortical nuclei and more severe seizures were observed in IC cortical microinjected animals. These audiogenic seizures were blocked with microinjections of 2-amino-7-phosphono-heptanoate (AP7) applied just before 2.5 μg NMDA microinjections into central or cortical nuclei. In S rats, AP7 totally blocked AS when microinjected into the central IC and partially, but significantly, blocked AS when applied into the cortical IC nucleus. In the last case, wild running was still present in 100% of the animals after AP7 treatment. These data may suggest an NMDA-dependent differential participation of IC subnuclei in the development of AS.

Neuroethological evaluation of audiogenic seizures and audiogenic-like seizures induced by microinjection of bicuculline into the inferior colliculus. II. Effects of nigral clobazam microinjections

Male Wistar rats were classified as susceptible (S) and resistant (R) to audiogenic seizures (AS) by evaluation of their response to high-intensity sound stimulation (110.3 dB). R rats injected with bicuculline into the inferior colliculus (IC) preferentially displayed audiogenic-like seizures with gyri, jumping and atonic falling, without important tonic-clonic components but with postictal contralateral asymmetry and hyperreactivity. These audiogenic-like seizures were blocked by clobazam microinjection into the substantia nigra (SN) and partially modified by SN vehicle injection. Injection of vehicle or clobazam into the SN of susceptible rats (S) did not modify the occurrence of AS. This may suggest the participation of GABAergic regulation in the development of audiogenic-like seizures in R rats and a defect in GABAergic neurotransmission in S rats.

The incidence and significance of the Tullio phenomenon in man.

The purpose of this study was to investigate the incidence and significance of the Tullio phenomenon in a group of human subjects. The subjects included 40 patients with complaints of auditory or vestibular symptoms. Ten otologically normal subjects were included in the study as a control group. All subjects underwent routine audiologic evaluation as well as electronystagmogram (ENG) testing. All subjects were then tested for the presence of the Tullio phenomenon by the method described. The results of this study showed that of the 40 subjects with known auditory or vestibular disorders, 90% (36) demonstrated nystagmus in response to high-intensity sound stimulation. All patients in the otologically normal control group demonstrated the presence of the Tullio phenomenon. No specific correlations were made between the presence of the Tullio phenomenon and specific audiologic or ENG findings. Studies on the effects of sound on the vestibular system are reviewed and lend support to the finding that the Tullio phenomenon may be a normal physiologic response in man under certain test conditions.