Cell-type-specific networks during hippocampal seizures at the micro- and macroscale

Abstract Epilepsy is a network disorder, involving neural circuits at both the micro- and macroscale. While local excitatory-inhibitory imbalances are recognized as a hallmark at the microscale, the dynamic role of distinct neuron types during seizures remain poorly understood. At the macroscale, interactions between key nodes within the epileptic network, such as the central median thalamic nucleus (CMT), are critical to the, hippocampal epileptic process. However, precise mechanisms underlying these interactions remain unclear. In this study, we investigated the microcircuit dynamics within the seizure onset zone and secondary spreading regions, as well as the network connectivity between the hippocampus and the CMT, using a 4-aminopyridine (4-AP) induced hippocampal seizure model. Rats were allocated into three experimental groups. The first group used a 3D tetrode array to monitor hippocampal seizure activity and microcircuit dynamics, including seizure propagation across the macroscale network. In the second group, a chemical lesion was induced in the CMT to assess its impact on hippocampal seizures. In the third group, chemogenetic techniques were used to selectively suppress pyramidal neurons in the CMT and observe changes in neural network connectivity between the CMT and hippocampus during seizures. Offline single-unit sorting was performed using KlustaKwik and further analysis was conducted with CellExplorer. At seizure onset, the narrow interneurons exhibited increased firing rates, initiating recruitment of other neurons, followed by increased activity in pyramidal neuron. Wide interneurons also showed heightened activity subsequent to pyramidal neurons. Interneurons played a more prominent role in the microcircuit during seizures compared to baseline. The CMT exhibited characteristic seizure activity and a decrease in narrow interneuron activity, whereas the cortex did not display seizure activity during hippocampal seizures. Lesioning the CMT resulted in the loss of the tonic component of hippocampal seizures and reduced overall neuronal activity in the hippocampal. Selective suppression of CMT pyramidal neurons resulted in shortened hippocampal seizures while preserving the tonic component. Narrow interneuron activity remained unchanged, while pyramidal neuron and wide interneuron activity significantly decreased. Our findings underscore the critical role of interneurons in the micronetwork of the seizure onset zone and secondary spreading region. Narrow interneurons were particularly vital in seizure initiation, whereas wide interneurons may contribute to seizure termination within the onset zone but not in the secondary spreading region. Pyramidal neurons in the CMT influence hippocampal seizures by modulating of both hippocampal pyramidal neurons and wide interneurons.