A pilot study of the role of the claustrum in attention and seizures in rats.

OBJECTIVE: The claustrum has been implicated in consciousness, and MRIs of patients with status epilepticus have shown increased claustral signal intensity. In an attempt to investigate the role of claustrum in cognition and seizures, we (1) assessed the effect of high-frequency stimulation (HFS) of the claustrum on performance in the operant chamber; (2) studied interclaustral and claustrohippocampal connectivity through cerebro-cerebral evoked potentials (CCEPs); and (3) investigated the role of claustrum in kainate-induced (KA) seizures. METHODS: Adult male Sprague-Dawley rats were trained in operant conditioning and implanted with electrodes in bilateral claustra and hippocampi. Claustrum HFS (50 Hz) was delivered bilaterally and unilaterally with increasing intensities from 50 to 1000 µA, and performance scores were assessed. CCEPs were studied by averaging the responses to bipolar stimulations, 1-ms wide pulses at 0.1 Hz to the claustrum. KA seizures were analyzed on video-EEG recordings. RESULTS: Generalized Estimating Equations analysis revealed that claustral stimulation reduced task performance scores relative to rest sessions (bilateral: -15.8 percentage points, p < 0.0001; unilateral: -15.2, p < 0.0001). With some stimulations, the rats showed a stimulus-locked decrease in attentiveness and, occasionally, an inability to complete the operant task. CCEPs demonstrated interclaustral and claustrohippocampal connectivity. Some KA seizures appeared to originate from the claustrum. CONCLUSIONS: Findings from the operant conditioning task suggest stimulation of the claustrum can alter attention or awareness. CCEPs demonstrated connectivity between the two claustra and between the claustrum and the hippocampi. Such connectivity may be part of the circuitry that underlies the alteration of awareness in limbic seizures. Lastly, KA seizures showed early involvement of the claustrum, a finding that also supports a possible role of the claustrum in the alteration of consciousness that accompanies dyscognitive seizures.

Effects of low-frequency electrical stimulation of the anterior piriform cortex on kainate-induced seizures in rats.

OBJECTIVE: Recent evidence in animals and humans suggests that low-frequency stimulation (LFS) has significant antiepileptic properties. The anterior piriform cortex (APC) is a highly susceptible seizure-trigger zone and may be critical for the initiation and propagation of seizures originating from cortical and limbic foci. We used the kainic acid (KA) seizure model in rats to assess the therapeutic effect of LFS of the APC on seizures. METHODS: Adult male Sprague-Dawley rats were implanted with electrodes in the left APC and recording electrodes bilaterally in the hippocampal CA3 regions. Rats were monitored continuously with video-EEG after the emergence of spontaneous recurrent seizures that followed induction of status epilepticus by intraperitoneal KA. After two weeks of baseline recordings to determine seizure frequency, LFS of the APC was applied 60-min On 15-min Off, for two weeks with 1Hz biphasic square waves, 0.2ms pulse width, at 200µA. Another 2-week period of video-EEG monitoring was done after the cessation of LFS to study the carry-over effect. Changes in seizure frequency, severity, and duration between baseline, during LFS, and post-LFS were analyzed using the Poisson regression model. RESULTS: Overall seizure frequency decreased during the post-LFS period to 5% of that at baseline (p=0.003). Severe seizures (stages 4 and 5 on the Racine scale) decreased to 0% of the baseline during the post-LFS period. CONCLUSIONS: Two weeks of LFS of the APC reduced spontaneous seizure frequency and severity in the KA model with the effect outlasting the stimulation. Our findings suggest that the APC can be an important therapeutic target for stimulation in epilepsy.

Spectral analysis of bilateral or alternate-site kindling-induced afterdischarges in the rabbit hippocampi.

Kindling is one of the popular animal models of temporal lobe epilepsy. In the present study following the previous results obtained using unilateral hippocampal kindling (UK), we performed spectral analysis of bilateral or alternate-site kindling-induced afterdischarges (ADs) in the rabbit hippocampi. Eight and ten adult rabbits were used for bilateral kindling (BK) and alternate-site kindling (AK), respectively. Kindling stimuli consisted of a train of biphasic pulses (1ms duration each) of 50Hz for 1s, with suprathreshold intensity for AD. The stimulations were applied simultaneously to the bilateral hippocampi in the BK and were delivered to the right and left hippocampus once every 24h in the AK. Motor responses were classified into five stages according to the conventional criteria. All animals in BK as well as AK developed stage 5 convulsions. This contrasts to the result of UK (kindled: 50%; incomplete: 50%). We normalized power spectral density (PSD) and monitored the changes in the proportion of lower frequency band component (LFB: 0-9Hz) and the higher frequency band (HFB: 12-30Hz). BK animals showed a significantly large decrement (0.5 times, p<0.01) in LFB component at the final stage compared to the initial stage, but a very large increment (4.7 times) in HFB component. Likewise, AK animals exhibited a significantly large decrement (0.6 times, p<0.01) in LFB component at the final stage, but a very large increment (3.6 times) in HFB component. Correlation analyses were performed between the HFB component and AD duration, interictal discharge frequency, and behavioral stages during kindling progression. Very strong positive correlations were found in both kindling animals. Chronological spectral analysis of seizure discharges, resulting in a pattern of LFB decrement accompanied by HFB increment, is a convenient tool to investigate epileptic disorders and diagnose epileptic states.

Fast Fourier transformation analysis of kindling-induced afterdischarge in the rabbit hippocampus.

Kindling is a widely used animal model of intractable temporal lobe epilepsy. In the present study, we performed fast Fourier transformation (FFT) analysis of kindling-induced afterdischarge (AD) in the rabbit hippocampus. Ten adult rabbits were used. Kindling stimulation to the right hippocampus was delivered as a train of biphasic pulses (1 ms duration each) of 50 Hz for 1s, with suprathreshold intensity for AD. Motor responses were classified into five stages according to the conventional criteria. Of 10 animals, five developed stage 5 convulsions with a mean of 21 stimulations (kindled (K) group), while the remaining five animals did not (incomplete kindling (IK) group). We standardized each ratio of power spectral density of lower frequency band component (LFB: 0-9 Hz) and the higher frequency band (HFB: 12-30 Hz) in the initial stage as 1.0. The IK group exhibited small decrements (0.99 and 0.94 times) in LFB and HFB components at the final stage. In contrast, the K group exhibited a significantly (p<0.05) large decrement (0.49 times) in the LFB component and a very large increment (4.45 times) of HFB component at the final stage. Correlation analyses were performed between alteration of power spectral density ratio of the HFB component and AD duration, interictal discharge frequency, and behavioral stage during kindling progression. Fairly strong positive correlations were found in all cases in the K group. FFT analysis of kindling-induced AD demonstrated an important role of the HFB component: enhancement of the HFB component is associated with kindled stage, while decrement of it is associated with incomplete kindling stage. These findings suggest that FFT analysis of stimulus-induced and spontaneous seizure discharges is useful for examination of the progression of epileptic disorders.