MicroPET Outperforms Beta-Microprobes in Determining Neuroreceptor Availability under Pharmacological Restriction for Cold Mass Occupancy.

Both non-invasive micro-positron emission tomography (µPET) and in situ beta-microprobes have the ability to determine radiotracer kinetics and neuroreceptor availability in vivo. Beta-microprobes were proposed as a cost-effective alternative to µPET, but literature revealed conflicting results most likely due to methodological differences and inflicted tissue damage. The current study has three main objectives: (i) evaluate the theoretical advantages of beta-microprobes; (ii) perform µPET imaging to assess the impact of (beta-micro)probe implantation on relative tracer delivery (R1) and receptor occupancy (non-displaceable binding potential, BPND) in the rat brain; and (iii) investigate whether beta-microprobe recordings produce robust results when a pharmacological restriction for cold mass dose (tracer dose condition) is imposed. We performed acquisitions (n = 61) in naive animals, dummy probe implanted animals (outer diameter: 0.75 and 1.00 mm) and beta-microprobe implanted animals (outer diameter: 0.75 mm) using two different radiotracers with high affinity for the striatum: [11C]raclopride (n = 29) and [11C]ABP688 (n = 32). In addition, acquisitions were completed with or without an imposed restriction for cold mass occupancy. We estimated BPND and R1 values using the simplified reference tissue method (SRTM). [11C]raclopride dummy µPET BPND (0.75 mm: -13.01 ± 0.94%; 1.00 mm: -13.89 ± 1.20%) and R1 values (0.75 mm: -29.67 ± 4.94%; 1.00 mm: -39.07 ± 3.17%) significantly decreased at the implant side vs. the contralateral intact side. A similar comparison for [11C]ABP688 dummy µPET, demonstrated significantly (p < 0.05) decreased BPND (-19.09 ± 2.45%) and R1 values (-38.12 ± 6.58%) in the striatum with a 1.00 mm implant, but not with a 0.75 mm implant. Particularly in tracer dose conditions, despite lower impact of partial volume effects, beta-microprobes proved unfit to produce representative results due to tissue destruction associated with probe insertion. We advise to use tracer dose µPET to obtain accurate results concerning receptor availability and tracer delivery, keeping in mind associated partial volume effects for which it is possible to correct.

Accelerated high-frequency repetitive transcranial magnetic stimulation enhances motor activity in rats.

High-frequency repetitive transcranial magnetic stimulation (HF-rTMS) is currently accepted as an evidence-based treatment option for treatment-resistant depression (TRD). Additionally, HF-rTMS showed beneficial effects on psychomotor retardation in patients. The classical HF-rTMS paradigms however are unlikely to replace electroconvulsive therapy, a more potent alternative for TRD albeit with important side-effects. Therefore, recent studies have investigated 'accelerated' HF-rTMS protocols demonstrating promising clinical responses in patients with TRD. Since the neuronal effects of accelerated HF-rTMS are underinvestigated, we evaluate here the possible metabolic and neurochemical effects of this treatment alternative. More specifically, we measured the effect on brain glucose metabolism and monoamines/metabolites, as well as on the spontaneous motor activity in rats. We found that brain glucose metabolism and monoamines remained generally unaffected after accelerated HF-rTMS, with the exception of reduced total striatal 5-hydroxyindolacetic acid (a metabolite of serotonin) levels. Interestingly, when compared to sham stimulation, the velocity, the total distance traveled as well as the percentage of movement, as measured by the open-field test, were significantly enhanced after accelerated HF-rTMS showing an increased motor activity. Our current results indicate that the accelerated HF-rTMS-induced increase in motor activity in rats, may be related to the striatal neurochemical effect.

Rat brain normalization templates for robust regional analysis of [11C]ABP688 positron emission tomography/computed tomography.

A methodology to generate rat brain templates for spatial normalization of positron emission tomographic (PET)/computed tomographic (CT) images is described and applied to generate three different templates for imaging of [11C]ABP688, a PET ligand binding to the metabotropic glutamate 5 receptor. The templates are based on functional (PET), structural (CT), and combined PET and CT information, respectively. The templates are created from a test-retest study under normal conditions and are used to assess the different templates by using them in the analysis pipeline of a test-retest and a blocking experiment. The resulting average nondisplaceable binding potentials (BPND) show significant (analysis of variance, p < .05) and substantial (up to 23%) differences between the different approaches in several brain regions. The highest BPND values in receptor-rich regions are obtained using the PET-based approach. This approach also had the smallest variability in all tested regions (standard error of measurement of 9% versus 14% [PET/CT] and 20% [CT]). All approaches showed similar relative changes in BPND values with increased blocking. Taken together, these results suggest that the use of the tracer-specific PET-based template outperforms the other approaches with the performance of the combined PET/CT template between those of the PET and the tracer-independent CT template.

The [18F]FDG µPET readout of a brain activation model to evaluate the metabotropic glutamate receptor 2 positive allosteric modulator JNJ-42153605.

Using [(18)F]fluorodeoxyglucose µ-positron emission tomography ([(18)F]FDG µPET), we compared subanesthetic doses of memantine and ketamine on their potential to induce increases in brain activation. We also studied the reversal effect of the well-known metabotropic glutamate receptor (mGluR)-2/3 agonist LY404039 [(-)-(1R,4S,5S,6S)-4-amino-2-sulfonylbicyclo[3.1.0]hexane-4,6-dicarboxylic acid] and the novel mGluR2 positive allosteric modulator (PAM) JNJ-42153605 [3-cylcopropylmethyl-7-(4-phenylpiperidin-1-yl)-8-trifluoromethyl [1,2,4] triazolo[4,3-a]pyridine]. First, rats (n = 12) were subjected to LY404039 (10 mg/kg s.c.) or vehicle, 30 minutes prior to saline, ketamine (30 mg/kg i.p.), or memantine (20 mg/kg i.p.). Second, rats (n = 12) were subjected to 2.5 mg/kg or 10 mg/kg mGluR2 PAM JNJ-42153605 or vehicle (s.c.), 30 minutes prior to memantine (20 mg/kg i.p.) or saline. Fifteen minutes later, [(18)F]FDG was injected (37 MBq i.v.) followed by a µPET/computed tomography scan. The increase due to memantine is significant for all relevant brain areas, whereas for ketamine this is not the case. Standard uptake values (SUVs) of the LY404039 pretreated and memantine-challenged group display a full reversal. Pretreatment with JNJ-42153605 also dose-dependently decreases SUV with a full reversal as well (for 10 mg/kg). Moreover, specificity of JNJ-42153605 is reached at this dose. In conclusion, this µPET experiment clearly indicates that subanesthetic doses of memantine induce significant increases of [(18)F]FDG SUVs in discrete brain areas and that the novel mGluR2 PAM has the capacity to dose-dependently and specifically reverse memantine-induced brain activation.

N-acetylcysteine- and MK-801-induced changes in glutamate levels do not affect in vivo binding of metabotropic glutamate 5 receptor radioligand 11C-ABP688 in rat brain.

UNLABELLED: Abnormal glutamate transmission is involved in various neurologic disorders, such as epilepsy, schizophrenia, and Parkinson disease. At present, no imaging techniques are capable of measuring acute fluctuations in endogenous glutamate levels in vivo. We evaluated the potential of (11)C-ABP688, a PET ligand that binds to an allosteric site of the metabotropic glutamate 5 receptor, in rats by using small-animal PET and ß-microprobes after pharmacologic challenges with N-acetylcysteine (NAc) and MK-801. Both compounds are known to induce increases in endogenous glutamate levels. METHODS: Three experiments with (11)C-ABP688 were performed to validate our study setup: first, metabolite analyses during workup (n = 3) and after a selected treatment (n = 3); second, a test-retest (n = 12) small-animal PET experiment (1 h scan; 27.75 MBq of (11)C-ABP688 administered intravenously; <3 nmol/kg); and third, a small-animal PET and ß-microprobe cold blocking study (n = 6/condition) with unlabeled ABP688. After this experimental validation, rats were pretreated with either NAc (intravenous infusion of 50 mg/kg/h) or MK-801 (0.16 mg/kg; given intraperitoneally); this step was followed by small-animal PET with (11)C-ABP688 (n = 12) or ß-microprobe measurements (n = 10/condition) of (11)C-ABP688. Time-activity curves were extracted, and the nondisplaceable binding potential (BPND) was calculated by use of the simplified reference tissue model with the cerebellum as a reference region. RESULTS: (11)C-ABP688 BPND measurements were highly reproducible (test-retest), and both small-animal PET and ß-microprobes were able to discriminate changes in (11)C-ABP688 binding (cold blocking). The average small-animal PET BPND measurements in the test experiment for the caudate putamen, frontal cortex, cerebral cortex, hippocampus, and thalamus were 2.58, 1.40, 1.60, 1.86, and 1.09, respectively. However, no significant differences in BPND measurements were observed with small-animal PET in the test and retest conditions on the one hand and the NAc and MK-801 conditions on the other hand for any of these regions. When ß-microprobes were used, the average BPND in the caudate putamen was 0.94, and no significant changes in the test and MK-801 conditions were observed. CONCLUSION: Pharmacologic challenges with NAc and MK-801 did not affect the (11)C-ABP688 BPND in the rat brain. These data suggest that the in vivo affinity of (11)C-ABP688 for binding to an allosteric site of the metabotropic glutamate 5 receptor is not modulated by changes in glutamate levels and that (11)C-ABP688 is not capable of measuring acute fluctuations in endogenous levels of glutamate in vivo in the rat brain.

Quantifying the effect of repetitive transcranial magnetic stimulation in the rat brain by µSPECT CBF scans.

BACKGROUND: Repetitive transcranial magnetic stimulation (rTMS) is used to treat neurological and psychiatric disorders such as depression and addiction amongst others. Neuro-imaging by means of SPECT is a non-invasive manner of evaluating regional cerebral blood flow (rCBF) changes, which are assumed to reflect changes in neural activity. OBJECTIVE: rCBF changes induced by rTMS are evaluated by comparing stimulation on/off in different stimulation paradigms using microSPECT of the rat brain. METHODS: Rats (n = 6) were injected with 10 mCi of (99m)Tc-HMPAO during application of two rTMS paradigms (1 Hz and 10 Hz, 1430 A at each wing of a 20 mm figure-of-eight coil) and sham. SPM- and VOI-based analysis was performed. RESULTS: rTMS caused widespread significant hypoperfusion throughout the entire rat brain. Differences in spatial extent and intensity of hypoperfusion were observed between both stimulation paradigms: 1 Hz caused significant hypoperfusion (P < 0.05) in 11.9% of rat brain volume while 10 Hz caused this in 23.5%; the minimal t-value induced by 1 Hz was -24.77 while this was -17.98 due to 10 Hz. Maximal percentage of hypoperfused volume due to 1 Hz and 10 Hz was reached at tissue experiencing 0.03-0.15 V/m. CONCLUSION: High-frequency (10 Hz) stimulation causes more widespread hypoperfusion, while 1 Hz induces more pronounced hypoperfusion. The effect of rTMS is highly dependent on the electric field strength in the brain tissue induced by the TMS coil. This innovative imaging approach can be used as a fast screening tool in quantifying and evaluating the effect of various stimulation paradigms and coil designs for TMS and offers a means for research and development.

Automatic detection of epileptic seizures on the intra-cranial electroencephalogram of rats using reservoir computing.

INTRODUCTION: In this paper we propose a technique based on reservoir computing (RC) to mark epileptic seizures on the intra-cranial electroencephalogram (EEG) of rats. RC is a recurrent neural networks training technique which has been shown to possess good generalization properties with limited training. MATERIALS: The system is evaluated on data containing two different seizure types: absence seizures from genetic absence epilepsy rats from Strasbourg (GAERS) and tonic-clonic seizures from kainate-induced temporal-lobe epilepsy rats. The dataset consists of 452hours from 23 GAERS and 982hours from 15 kainate-induced temporal-lobe epilepsy rats. METHODS: During the preprocessing stage, several features are extracted from the EEG. A feature selection algorithm selects the best features, which are then presented as input to the RC-based classification algorithm. To classify the output of this algorithm a two-threshold technique is used. This technique is compared with other state-of-the-art techniques. RESULTS: A balanced error rate (BER) of 3.7% and 3.5% was achieved on the data from GAERS and kainate rats, respectively. This resulted in a sensitivity of 96% and 94% and a specificity of 96% and 99% respectively. The state-of-the-art technique for GAERS achieved a BER of 4%, whereas the best technique to detect tonic-clonic seizures achieved a BER of 16%. CONCLUSION: Our method outperforms up-to-date techniques and only a few parameters need to be optimized on a limited training set. It is therefore suited as an automatic aid for epilepsy researchers and is able to eliminate the tedious manual review and annotation of EEG.

Repeated assessment of larynx compound muscle action potentials using a self-sizing cuff electrode around the vagus nerve in experimental rats.

RATIONALE: Vagus nerve stimulation (VNS) is an adjunctive treatment for patients with refractory epilepsy. In more than 30% of the patients VNS has no therapeutic effect. The goal of this study was to find an objective parameter that can be used as an indicator of effective stimulation of the vagus nerve. METHODS: The electrophysiological response to VNS was recorded from the vagus nerve, recurrent laryngeal nerve and larynx muscles. Nerve lesions and muscle relaxing agent were used to find the source of the electrophysiological response. A cuff-electrode for chronic stimulation and recording was implanted for chronic recording of the VNS-induced electrophysiological response after implantation. Dose-response curves were determined daily during a follow-up period of 2 months. RESULTS: VNS induced an electrophysiological response around 3 ms after start of the stimulation. This response was identified as a larynx compound action potential (LCMAP) LCMAP could be recorded immediately after surgery in 11/21 rats, while in the other 10/21 rats, a recovery period with an average of 25 days was required. Once the LCAMP could be recorded, the latency and overall characteristics of the doses response curves of the LCMAP remained stable during the entire follow-up period. CONCLUSIONS: In this study, we provide an objective electrophysiological parameter for vagus nerve activation. LCAMP may indicate recovery of the vagus nerve after implantation, which may help to determine when uptitration of VNS therapy can be initiated. LCAMP could be of value in future experiments for objectification of VNS in animal models for epilepsy.

Increased hippocampal noradrenaline is a biomarker for efficacy of vagus nerve stimulation in a limbic seizure model.

Vagus nerve stimulation (VNS) is an effective adjunctive treatment for medically refractory epilepsy. In this study, we measured VNS-induced changes in hippocampal neurotransmitter levels and determined their potential involvement in the anticonvulsive action of VNS, to elucidate the mechanism of action responsible for the seizure suppressing effect of VNS in an animal model for limbic seizures. We used in vivo intracerebral microdialysis to measure VNS-induced changes in hippocampal extracellular concentrations of noradrenaline, dopamine, serotonin and GABA in freely moving, male Wistar rats. During the same experiment, the effect of VNS on pilocarpine-induced limbic seizures was assessed using video-EEG monitoring. The involvement of VNS-induced increases in hippocampal noradrenaline in the mechanims of action of VNS was evaluated by blocking hippocampal α(2)-receptors. VNS produced a significant increase in hippocampal noradrenaline concentration (69 ± 16% above baseline levels). VNS also increased the latency between pilocarpine infusion and the onset of epileptiform discharges, and reduced the duration and severity of pilocarpine-induced limbic seizures. A strong positive correlation was found between the noradrenergic and anticonvulsive effects of VNS. Blockade of hippocampal α(2 -receptors reversed the seizure-suppressing effect of VNS. VNS induces increases in extracellular hippocampal noradrenaline, which are at least partly responsible for its seizure-suppressing effect in a model for limbic seizures, and constitute a potential biomarker for the efficacy of VNS in temporal lobe epilepsy.

A novel implantable vagus nerve stimulation system (ADNS-300) for combined stimulation and recording of the vagus nerve: pilot trial at Ghent University Hospital.

PURPOSE: Vagus nerve stimulation (VNS) is an established treatment for refractory epilepsy. The ADNS-300 is a new system for VNS that includes a rechargeable stimulus generator and an electrode for combined stimulation and recording. In this feasibility study, three patients were implanted with ADNS-300 for therapeutic VNS. In addition, compound action potentials (CAPs) were recorded to evaluate activation of the vagus nerve in response to VNS. METHODS: Three patients were implanted with a cuff-electrode around the left vagus nerve, that was connected to a rechargeable pulse generator under the left clavicula. Two weeks after surgery, therapeutic VNS (0.25-1.25 mA, 500 µs, 30s on, 10 min off and 30Hz) was initiated and stimulus-induced CAPs were recorded. RESULTS: The ADNS-300 system was successfully implanted in all three patients and patients were appropriately stimulated during six months of follow-up. A reduction in seizure frequency was demonstrated in two patients (43% and 40% in patients 1 and 3, respectively), while in patient 2 seizure frequency remained unchanged. CAPs could be recorded in patients 1 and 2, proving stimulation-induced activation of the vagus nerve. CONCLUSION: This feasibility study demonstrates that the ADNS-300 system can be used for combined therapeutic stimulation (in 3/3 patients) and recording of CAPs in response to VNS (in 2/3 patients) up to three weeks after surgery. Implantation in a larger number of patients will lead to a better understanding of the electrophysiology of the vagus nerve, which in turn could result in more adequate and individualized VNS parameter choice.

Suppression of hippocampal epileptic seizures in the kainate rat by Poisson distributed stimulation.

PURPOSE: Hippocampal deep brain stimulation (DBS) is an experimental therapy for patients with pharmacoresistant temporal lobe epilepsy (TLE). Despite the successful clinical application of DBS, the optimal stimulation parameters are undetermined. We evaluate the efficacy of a new form of DBS, using continuous stimuli with Poisson distributed intervals (Poisson distributed stimulation, PDS) in the kainate (KA) rat model, a validated model for human TLE. METHODS: Status epilepticus was elicited by injection of KA (i.p.). After development of spontaneous seizures, rats were implanted with hippocampal DBS- and depth electroencephalography (EEG) electrodes. After baseline EEG monitoring, one group of rats (n = 13) was treated with PDS and a second (n = 11) received regular high frequency stimulation (HFS) at 130 Hz. Stimulation intensity was 100 µA below the threshold for induction of epileptiform EEG activity. RESULTS: Stimulation intensity was significantly lower for PDS (156 ± 20 µA) than HFS (207 ± 23 µA; p < 0.02). Seven (54%) of 13 rats treated with PDS and 5 (45%) of 11 rats treated with HFS experienced a significant reduction in seizure frequency. In PDS-improved rats, seizure frequency was reduced to 33% (p < 0.01) of baseline value and in HFS-improved rats to 50% (p < 0.01). After termination of PDS, seizure rate returned to baseline value. DISCUSSION: Continuous hippocampal PDS significantly reduces the number of spontaneous seizures. Compared to regular HFS, there is a slightly larger number of improved rats and a larger efficacy at a considerably lower stimulus intensity. The first two observations leave room for optimization, whereas a lower intensity is beneficial for battery life.

Continuous local intrahippocampal delivery of adenosine reduces seizure frequency in rats with spontaneous seizures.

PURPOSE: Despite different treatment options for patients with refractory epilepsy such as epilepsy surgery and neurostimulation, many patients still have seizures and/or drug-related cerebral and systemic side effects. Local intracerebral delivery of antiepileptic compounds may represent a novel strategy with specific advantages such as the option of higher local doses and reduced side effects. In this study we evaluate the antiepileptic effect of local delivery of adenosine in the kainic acid rat model, a validated model for temporal lobe epilepsy. METHODS: Fifteen rats, in which intraperitoneal kainic acid injection had induced spontaneous seizures, were implanted with a combination of depth electrodes and a cannula in both hippocampi. Cannulas were connected to osmotic minipumps to allow continuous hippocampal delivery. Rats were freely moving and permanently monitored by video-EEG (electroencephalography). Seizures were scored during 2 weeks of local hippocampal delivery of saline (baseline), followed by 2 weeks of local adenosine (6 mg/ml) (n = 10) or saline (n = 5) delivery (0.23 µl/h) (treatment). In 7 of 10 adenosine-treated rats, saline was also delivered during a washout period. RESULTS: During the treatment period a mean daily seizure frequency reduction of 33% compared to the baseline rate was found in adenosine-treated rats (p < 0.01). Four rats had a seizure frequency reduction of at least 50%. Both nonconvulsive and convulsive seizures significantly decreased during the treatment period. In the saline-control group, mean daily seizure frequency increased with 35% during the treatment period. CONCLUSIONS: This study demonstrates the antiseizure effect of continuous adenosine delivery in the hippocampi in rats with spontaneous seizures.

Hippocampal deep brain stimulation induces decreased rCBF in the hippocampal formation of the rat.

Deep brain stimulation (DBS) is a promising experimental approach to treat various neurological disorders. However, the optimal stimulation paradigm and the precise mechanism of action of DBS are unknown. Neuro-imaging by means of Single Photon Emission Computed Tomography (SPECT) is a non-invasive manner of evaluating regional cerebral blood flow (rCBF) changes, which are assumed to reflect changes in neural activity. In this study, rCBF changes induced by hippocampal DBS are evaluated by subtraction analysis of stimulation on/off using small animal microSPECT of the rat brain. Rats (n=13) were implanted with a multi-contact DBS electrode in the right hippocampus and injected with 10 mCi of HMPAO-Tc99(m) during application of various hippocampal DBS paradigms and amplitudes and during sham stimulation. Subtraction analysis revealed that hippocampal DBS caused a significant decrease in relative rCBF, both in the ipsi- (the side of the implanted electrode) and contralateral hippocampus. Hypoperfusion spread contralaterally with increasing stimulation amplitude. A clear distinction in spatial extent and intensity of hypoperfusion was observed between stimulation paradigms: bipolar Poisson Distributed Stimulation induced significant hypoperfusion ipsi- and contralaterally (p<0.01), while during other stimulation paradigms, rCBF-changes were less prominent. In conclusion, small animal microSPECT allows us to draw conclusions on the location, spatial extent and intensity of the hypoperfusion observed in the ipsi- and contralateral hippocampus, induced by hippocampal DBS. Our study demonstrates an innovative approach to visualize the effects of DBS and can be a useful tool in evaluating the effect of various stimulation paradigms and target areas for DBS.

Comparison of hippocampal Deep Brain Stimulation with high (130Hz) and low frequency (5Hz) on afterdischarges in kindled rats.

Hippocampal Deep Brain Stimulation (DBS) is proposed as an experimental treatment for refractory epilepsy, but the optimal stimulation parameters are undetermined. High frequency hippocampal DBS at 130Hz is effective in both animals and patients with epilepsy. Low frequency stimulation (approximately 5Hz) is assumed to have anti-epileptic properties but the efficacy is highly debated. This animal study compares the effects of both stimulation modalities in kindled rats. Sprague Dawley rats (n=20) were fully kindled according to the Alternate Day Rapid Kindling-protocol. After a baseline kindling period, rats were divided into a high frequency group (HFS, 130Hz, n=11) and a low frequency group (LFS, 5Hz, n=9), both receiving 10 days of continuous DBS. During and after DBS, the seizure susceptibility of all rats was tested and the characteristics of the afterdischarges (ADs) were compared between both treatments. During HFS, AD threshold was higher (p<0.05) and at the stimulated site, AD latency was longer (p<0.01) than during baseline period. During LFS, a similar but smaller change was observed, but did not reach significance. The duration of the AD was not affected by either HFS or LFS. After termination of HFS, the effects on AD latency and AD threshold recovered to baseline. In conclusion, high frequency stimulation at 130Hz is more effective than LFS (5Hz) in affecting excitability in epileptic rats. This is reflected in a higher AD threshold and longer AD latency during application of stimulation.

Unconditioned adult-derived neurosphere cells mainly differentiate towards astrocytes upon transplantation in sclerotic rat hippocampus.

PURPOSE: Cell transplantation is being investigated as an alternative treatment for medically refractory temporal lobe epilepsy (TLE). In this study the fate of adult-derived neurosphere cells was evaluated after transplantation in the lesioned hippocampus of the intrahippocampal kainic acid (KA) model for TLE. METHODS: Neurosphere-forming cells were derived from the subventricular zone (SVZ) of transgenic green fluorescent protein (GFP) reporter mice and expanded in culture. After 10 passages in vitro neurosphere-derived cells were transplanted in the hippocampus three days (KA3d group) and three weeks (KA3w group) after intrahippocampal KA injection. Survival and differentiation of neurosphere cells were evaluated three and six weeks after transplantation. RESULTS: A fraction (about 1%) of GFP-expressing neurosphere cells survived for at least six weeks after transplantation with a higher and more robust survival rate in the KA3d compared to the KA3w group. Although a small fraction of the cells expressed the neuronal marker NeuN, neurosphere cells mainly differentiated towards astrocytes. DISCUSSION: Our results indicate that adult-derived neurosphere cells are able to survive upon transplantation in the sclerotic hippocampus. The transplanted cells do not or hardly contribute to neuronal replacement and mainly adopt an astrogliotic fate.

Electrical stimulation for the treatment of epilepsy.

Despite the advent of new pharmacological treatments and the high success rate of many surgical treatments for epilepsy, a substantial number of patients either do not become seizure-free or they experience major adverse events (or both). Neurostimulation-based treatments have gained considerable interest in the last decade. Vagus nerve stimulation (VNS) is an alternative treatment for patients with medically refractory epilepsy, who are unsuitable candidates for conventional epilepsy surgery, or who have had such surgery without optimal outcome. Although responder identification studies are lacking, long-term VNS studies show response rates between 40% and 50% and long-term seizure freedom in 5% to 10% of patients. Surgical complications and perioperative morbidity are low. Research into the mechanism of action of VNS has revealed a crucial role for the thalamus and cortical areas that are important in the epileptogenic process. Acute deep brain stimulation (DBS) in various thalamic nuclei and medial temporal lobe structures has recently been shown to be efficacious in small pilot studies. There is little evidence-based information on rational targets and stimulation parameters. Amygdalohippocampal DBS has yielded a significant decrease of seizure counts and interictal EEG abnormalities during long-term follow-up. Data from pilot studies suggest that chronic DBS for epilepsy may be a feasible, effective, and safe procedure. Further trials with larger patient populations and with controlled, randomized, and closed-loop designs should now be initiated. Further progress in understanding the mechanism of action of DBS for epilepsy is a necessary step to making this therapy more efficacious and established.

High frequency deep brain stimulation in the hippocampus modifies seizure characteristics in kindled rats.

PURPOSE: This experimental animal study evaluates the effect of high frequency deep brain stimulation (HFS DBS) on seizures in the Alternate Day Rapid Kindling model for temporal lobe epilepsy (TLE). The target for HFS is the hippocampus, as this structure is often presumed to be the seizure focus in human TLE. METHODS: Rats (n = 12) were fully kindled in the hippocampus according to the Alternate Day Rapid Kindling protocol. Characteristics of the evoked afterdischarges (AD) were determined in the baseline period using AD threshold, AD latency, and AD duration as parameters. Rats were divided into a treated group (n = 7) that received 130 Hz HFS for 1 week, and a control group (n = 5) that did not receive HFS. Rats were retested in the following week. After 1 additional week of rest, the HFS group was continuously stimulated again for 1 week, during which AD evoked by kindling stimuli were characterized again. RESULTS: HFS had a direct effect on evoked AD: during HFS, it increased AD threshold to 203 +/- 13% of controls (p < 0.01) and increased AD latency to 191 +/- 19% (p < 0.05). It decreased AD duration to 71 +/- 9% (p < 0.05) of controls. The effect outlasted the HFS stimulation as in the week following HFS similar differences, but smaller in size, could still be established. CONCLUSION: Continuous HFS (130 Hz) in the hippocampus of epileptic rats modulates the characteristics of evoked AD in a way that reflects a reduction in excitability of the target region.

Neuromodulation with levetiracetam and vagus nerve stimulation in experimental animal models of epilepsy.

Epilepsy is a neurological disorder consisting of recurrent seizures, resulting from excessive, uncontrolled electrical activity in the brain. Epilepsy treatment is successful in the majority of the cases; however; still one third of the epilepsy patients are refractory to treatment. Besides the ongoing research on the efficacy of antiepileptic treatments in suppressing seizures (anti-seizure effect), we want to seek for therapies that can lead to plastic, neuromodulatory changes in the epileptic network. Neuropharmacological therapy with levetiracetam (LEV) and vagus nerve stimulation (VNS) are two novel treatments for refractory epilepsy. LEV acts rapidly on seizures in both animal models and humans. In addition, preclinical studies suggest that LEV may have antiepileptogenic and neuroprotective effects, with the potential to slow or arrest disease progression. VNS as well can have an immediate effect on seizures in epilepsy models and patients with, in addition, a cumulative effect after prolonged treatment. Studies in man are hampered by the heterogeneity of patient populations and the difficulty to study therapy-related effects in a systematic way. Therefore, investigation was performed utilizing two rodent models mimicking epilepsy in humans. Genetic absence epilepsy rats from Strasbourg (GAERS) have inborn absence epilepsy and Fast rats have a genetically determined sensitivity for electrical amygdala kindling, which is an excellent model of temporal lobe epilepsy. Our findings support the hypothesis that treatment with LEV and VNS can be considered as neuromodulatory: changes are induced in central nervous system function or organization as a result of influencing and initiating neurophysiological signals.

Rapid kindling in preclinical anti-epileptic drug development: the effect of levetiracetam.

PURPOSE: This study assesses the use of the serial day Rapid Kindling with Recurrent Hippocampal Seizures (RKRHS) model in drug testing by investigating the anti-epileptic effect of levetiracetam (LEV), a novel anti-epileptic drug (AED) with a unique preclinical profile. METHODS: Male Wistar rats (n=16) were implanted with a stimulation/recording electrode unit in the right hippocampus and epidural recording electrodes. One week later, all rats received 12 stimulations each day for several consecutive days according to the serial day RKRHS protocol, until they were fully kindled. Fully kindled rats were then randomly assigned to an active (n=8) and a control (n=6) group and injected once (intraperitoneal, i.p.) with levetiracetam (54 mg/kg) or saline (0.9% NaCl, 2 ml/kg), respectively. One hour later, the rats received additional kindling stimulations, during which the effect of LEV was assessed. RESULTS: One hour following injection of LEV, mean seizure stage dropped to 1.67+/-1.03 compared to 5+/-0 in controls (p<0.05). Mean ADD was also significantly shorter in the active group than in controls; 21.16+/-5.03 s versus 57.24+/-8.16s, respectively (p<0.05). A gradual, time-dependent decline was noted in the anti-epileptic effect of LEV but this effect stayed statistically significant at least up to 2.5 h (p<0.05). CONCLUSION: LEV was demonstrated to have anti-epileptic properties in RKRHS that compared to those in traditional kindling and contrast with results from classical screening tests. RKRHS may represent a valuable and sensitive screening tool early in the drug screening process.