Chemogenetic activation of hippocampal area CA2 promotes acute and chronic seizures in a mouse model of epilepsy

Neurobiology of Disease · 2026-01-03 · 1 citations

Pyramidal cells (PCs) of hippocampal area CA2 exhibit increased excitability in temporal lobe epilepsy (TLE) and in mouse models of TLE. In epileptic mice, selective inhibition of CA2 PCs reduces chronic seizures. Here we asked if activating CA2 PCs increases seizures. Mice expressing Cre recombinase in CA2 PCs (Amigo2-Cre mice) were injected with the convulsant pilocarpine to induce a period of severe seizures (status epilepticus, SE), which leads to chronic seizures after 3-4 weeks (epilepsy). Epileptic mice were injected with a Cre-dependent adeno-associated virus (AAV) to express an excitatory designer receptor exclusively activated by designer drug (eDREADD; hM3Dq) in dorsal CA2 bilaterally and implanted with subdural EEG electrodes. After recovery, mice were recorded continuously using video and EEG for 6 weeks, 3 weeks with drinking water containing the eDREADD activator clozapine-N-oxide (CNO) and 3 weeks without CNO. CA2 activation with CNO caused a significant increase in seizure frequency and duration. Seizures occurred in clusters (many seizures per day over several consecutive days) and mice given water with CNO had a greater maximum number of seizures per day during a cluster compared to water without CNO. CNO had no significant effect in control mice. In naïve Amigo2-Cre mice expressing hM3Dq, pre-treatment with CNO before pilocarpine administration shortened the latency to SE and increased EEG power at the start of SE. Taken together with prior findings, the results suggest that CA2 is a control point for regulating seizures in the pilocarpine mouse model of TLE.

Chemogenetic activation of hippocampal area CA2 promotes acute and chronic seizures in a mouse model of epilepsy

bioRxiv (Cold Spring Harbor Laboratory) · 2025-12-29

ABSTRACT Pyramidal cells (PCs) of hippocampal area CA2 exhibit increased excitability in temporal lobe epilepsy (TLE) and in mouse models of TLE. In epileptic mice, selective inhibition of CA2 PCs reduces chronic seizures. Here we asked if activating CA2 PCs increases seizures. Mice expressing Cre recombinase in CA2 PCs ( Amigo2 -Cre mice) were injected with the convulsant pilocarpine to induce a period of severe seizures ( status epilepticus , SE), which leads to chronic seizures after 3-4 weeks (epilepsy). Epileptic mice were injected with a Cre-dependent adeno-associated virus (AAV) to express an excitatory designer receptor exclusively activated by designer drug (eDREADD; hM3Dq) in dorsal CA2 bilaterally and implanted with subdural EEG electrodes. After recovery, mice were recorded continuously using video and EEG for 6 weeks, 3 weeks with drinking water containing the eDREADD activator clozapine-N-oxide (CNO) and 3 weeks without CNO. CA2 activation with CNO caused a significant increase in seizure frequency and duration. Seizures occurred in clusters (many seizures per day over several consecutive days) and mice given water with CNO had a greater maximum number of seizures per day during a cluster compared to water without CNO. CNO had no significant effect in control mice. In naïve Amigo2- Cre mice expressing hM3Dq, pre-treatment with CNO before pilocarpine administration shortened the latency to SE and increased EEG power at the start of SE. Taken together with prior findings, the results suggest that CA2 is a control point for regulating seizures in the pilocarpine mouse model of TLE. HIGHLIGHTS Chemogenetic excitation of CA2 increased chronic seizure frequency and duration in epileptic mice. When seizures occurred in clusters, CA2 excitation increased the peak of the cluster. When sexes were separated, males showed effects on seizure frequency but both sexes showed effects on seizure duration and clusters. Chemogenetic excitation of CA2 promoted pilocarpine-induced status epilepticus in normal mice. GRAPHICAL ABSTRACT Legend: In adult mice, injecting the convulsant pilocarpine induces severe seizures ( status epilepticus , SE) and ultimately leads to chronic intermittent seizures (epilepsy). Selective activation of hippocampal area CA2 during the period of chronic seizures led to greater seizure frequency and duration, as well as a greater number of seizures/day at the peak of a seizure cluster. Effects were greatest in males. In naive mice, activation of CA2 prior to pilocarpine administration facilitated SE. Therefore, CA2 influences acute and chronic seizures in this mouse model.

Tuned geometries of hippocampal representations meet the computational demands of social memory

Neuron · 2024-02-20 · 71 citations

Vivien Chevaleyre (1973–2024)

Neuron · 2024-05-01

Regulation by Presynaptic NMDA Receptors of Non-Linear Postsynaptic Summation of the Cortical Input to CA1 Pyramidal Neurons

Neuroscience · 2024-06-13

Modulation of Aggression by Social Novelty Recognition Memory in the Hippocampal CA2 Region

SSRN Electronic Journal · 2024-01-01 · 1 citations

A neural mechanism for discriminating social threat from social safety

bioRxiv (Cold Spring Harbor Laboratory) · 2023-07-04 · 2 citations

The ability to distinguish a threatening from non-threatening conspecific based on past experience is critical for adaptive social behaviors. Although recent progress has been made in identifying the neural circuits that contribute to different types of positive and negative social interactions, the neural mechanisms that enable the discrimination of individuals based on past aversive experiences remain unknown. Here, we developed a modified social fear conditioning paradigm that induced in both sexes robust behavioral discrimination of a conspecific associated with a footshock (CS+) from a non-reinforced interaction partner (CS-). Strikingly, chemogenetic or optogenetic silencing of hippocampal CA2 pyramidal neurons, which have been previously implicated in social novelty recognition memory, resulted in generalized avoidance fear behavior towards the equally familiar CS-and CS+. One-photon calcium imaging revealed that the accuracy with which CA2 representations discriminate the CS+ from the CS-animal was enhanced following social fear conditioning and strongly correlated with behavioral discrimination. Moreover the CA2 representations incorporated a generalized or abstract representation of social valence irrespective of conspecific identity and location. Thus, our results demonstrate, for the first time, that the same hippocampal CA2 subregion mediates social memories based on conspecific familiarity and social threat, through the incorporation of a representation of social valence into an initial representation of social identity.

Reduced Cholecystokinin-Expressing Interneuron Input Contributes to Disinhibition of the Hippocampal CA2 Region in a Mouse Model of Temporal Lobe Epilepsy

Journal of Neuroscience · 2023-08-29 · 13 citations

A significant proportion of temporal lobe epilepsy (TLE) patients experience drug-resistant seizures associated with mesial temporal sclerosis, in which there is extensive cell loss in the hippocampal CA1 and CA3 subfields, with a relative sparing of dentate gyrus granule cells and CA2 pyramidal neurons (PNs). A role for CA2 in seizure generation was suggested based on findings of a reduction in CA2 synaptic inhibition (Williamson and Spencer, 1994) and the presence of interictal-like spike activity in CA2 in resected hippocampal tissue from TLE patients (Wittner et al., 2009). We recently found that in the pilocarpine-induced status epilepticus (PILO-SE) mouse model of TLE there was an increase in CA2 intrinsic excitability associated with a loss of CA2 synaptic inhibition. Furthermore, chemogenetic silencing of CA2 significantly reduced seizure frequency, consistent with a role of CA2 in promoting seizure generation and/or propagation (Whitebirch et al., 2022). In the present study, we explored the cellular basis of this inhibitory deficit using immunohistochemical and electrophysiological approaches in PILO-SE male and female mice. We report a widespread decrease in the density of pro-cholecystokinin-immunopositive (CCK + ) interneurons and a functional impairment of CCK + interneuron-mediated inhibition of CA2 PNs. We also found a disruption in the perisomatic perineuronal net in the CA2 stratum pyramidale. Such pathologic alterations may contribute to an enhanced excitation of CA2 PNs and CA2-dependent seizure activity in the PILO-SE mouse model. SIGNIFICANCE STATEMENT Impaired synaptic inhibition in hippocampal circuits has been identified as a key feature that contributes to the emergence and propagation of seizure activity in human patients and animal models of temporal lobe epilepsy (TLE). Among the hippocampal subfields, the CA2 region is particularly resilient to seizure-associated neurodegeneration and has been suggested to play a key role in seizure activity in TLE. Here we report that perisomatic inhibition of CA2 pyramidal neurons mediated by cholecystokinin-expressing interneurons is selectively reduced in acute hippocampal slices from epileptic mice. Parvalbumin-expressing interneurons, in contrast, appear relatively conserved in epileptic mice. These findings advance our understanding of the cellular mechanisms underlying inhibitory disruption in hippocampal circuits in a mouse model of spontaneous recurring seizures.

Hippocampal area CA2 controls seizure dynamics, interictal EEG abnormalities and social comorbidity in mouse models of temporal lobe epilepsy

bioRxiv (Cold Spring Harbor Laboratory) · 2023-01-19 · 15 citations

Temporal lobe epilepsy (TLE) is characterized by spontaneous recurrent seizures, abnormal activity between seizures, and impaired behavior. CA2 pyramidal neurons (PNs) are potentially important because inhibiting them with a chemogenetic approach reduces seizure frequency in a mouse model of TLE. However, whether seizures could be stopped by timing inhibition just as a seizure begins is unclear. Furthermore, whether inhibition would reduce the cortical and motor manifestations of seizures are not clear. Finally, whether interictal EEG abnormalities and TLE comorbidities would be improved are unknown. Therefore, real-time optogenetic silencing of CA2 PNs during seizures, interictal activity and behavior were studied in 2 mouse models of TLE. CA2 silencing significantly reduced seizure duration and time spent in convulsive behavior. Interictal spikes and high frequency oscillations were significantly reduced, and social behavior was improved. Therefore, brief focal silencing of CA2 PNs reduces seizures, their propagation, and convulsive manifestations, improves interictal EEG, and ameliorates social comorbidities. HIGHLIGHTS: Real-time CA2 silencing at the onset of seizures reduces seizure durationWhen CA2 silencing reduces seizure activity in hippocampus it also reduces cortical seizure activity and convulsive manifestations of seizuresInterictal spikes and high frequency oscillations are reduced by real-time CA2 silencingReal-time CA2 silencing of high frequency oscillations (>250Hz) rescues social memory deficits of chronic epileptic mice.

Lipophilic Tracer DiI Used for Neuronal Tracing in the Fixed Hippocampal Formations of Mice

Columbia Undergraduate Science Journal · 2022-03-29

Both the hippocampus and the entorhinal cortex are involved in memory formation. It is thought that they work together to create a loop that is involved in the formation of long term memories. The entorhinal cortex sends projections to the CA1 field of the hippocampus and the CA1 field sends projections back to the entorhinal cortex. How exactly they communicate is unknown. In order to do electrophysiological studies it is necessary to know in which orientation to cut the brain to preserve the most connections between these two areas. Here we show that DiI crystal application can be used to successfully label known fiber paths from the dentate gyrus to the CA3 field. Then we begin with sagittal and horizontal slices of the brain to see how well connections between the CA1 field and the EC are preserved.