Abstract
BackgroundHippocampal and cortical slice-based models are widely used to study seizures and epilepsy. Seizure detection and quantification are essential components for studying mechanisms of epilepsy and assessing therapeutic interventions. To obtain meaningful signals and maximize experimental throughput, variability should be minimized. Some electrical recording methods require insertion of an electrode into neuronal tissue, change in slice chemical microenvironment, and transients in temperature and pH. These perturbations can cause acute and long-term alterations of the neuronal network which may be reflected in the variability of the recorded signal. New methodIn this study we investigated the effect of experimental perturbations in three local field potential (LFP) recording methods including substrate micro-wires (s-MWs), multiple electrode arrays (MEAs), and inserted micro wire electrodes (i-MW). These methods enabled us to isolate effects of different perturbations. We used organotypic hippocampal slices (OHCs) as an in-vitro model of posttraumatic epilepsy. To investigate the effect of the disturbances caused by the recording method on the paroxysmal events, we introduced jitter analysis, which is sensitive to small differences in the seizure spike timing. ResultsMedium replacement can introduce long-lasting perturbations. Electrode insertion increased variability on a shorter time scale. OHCs also underwent spontaneous state transitions characterized by transient increases in variability. Comparison with existing methodsThis new method of seizure waveform analysis allows for more sensitive assessment of variability of ictal events than simply measuring seizure frequency and duration. ConclusionWe demonstrated that some of the variability in OHC recordings are due to experimental perturbations while some are spontaneous and independent of recording method.
Highlights
Hippocampal and cortical slice-based models are widely used to study seizures and epilepsy (Pitk€anen et al, 2017)
In order to record from substrate micro-wires (s-MWs), culture plates were transferred to the recording chamber, and to record from cultures on coverslips through the inserted micro wire electrodes (i-MW), the coverslip was transferred to a recording interface
In all three recording modalities (s-MW, i-MW, and multiple electrode arrays (MEAs)) large-area, low-impedance reference electrodes were used
Summary
Hippocampal and cortical slice-based models are widely used to study seizures and epilepsy (Pitk€anen et al, 2017). Some electrical recording methods require insertion of an electrode into neuronal tissue, change in slice chemical microenvironment, and transients in temperature and pH These perturbations can cause acute and long-term alterations of the neuronal network which may be reflected in the variability of the recorded signal. New method: In this study we investigated the effect of experimental perturbations in three local field potential (LFP) recording methods including substrate micro-wires (s-MWs), multiple electrode arrays (MEAs), and inserted micro wire electrodes (i-MW). These methods enabled us to isolate effects of different perturbations. Conclusion: We demonstrated that some of the variability in OHC recordings are due to experimental perturbations while some are spontaneous and independent of recording method
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