C1q neutralization during epileptogenesis attenuates complement-mediated synaptic elimination and epileptiform...

Accumulating evidence indicates that aberrant C1q-C3 complement signaling in microglia and astrocytes drives synaptic dysfunction and neuronal loss. C1q-mediated synaptic dysfunction disrupts neuronal circuitry balance and can lead to network hyperexcitability in epilepsy. However, the therapeutic potential of C1q inhibition remains unclear. This study aimed to delineate whether blocking C1q-driven pathways at critical times can effectively prevent further synaptic loss and attenuate epileptiform activity in a mouse model of epilepsy.

Microglial C1q and astrocytic C3 expression was examined in hippocampal CA1 and CA3 regions at acute (1 day), subacute (7, 14 days), and chronic (5 weeks) phases following pilocarpine-induced status epilepticus (SE) in mice. To evaluate the role of C1q activation, a C1q-neutralizing antibody or isotype control immunoglobulin G antibody was administered into cisterna magna at 7 days post-SE. During the chronic phase, glial reactivity, dendritic spine density, and epileptiform activity were assessed by immunostaining, Golgi staining, and electroencephalography (EEG) recording. After exclusions, 58 mice were included in the study.

C1q expression was elevated during the subacute phase (7 and 14 days post-SE) in CA1 and CA3 regions; astrocytic C3 expression increased in CA1 and persisted chronically. C1q-neutralizing antibody administration at 7 days post-SE, a time point characterized by prominent C1q upregulation, attenuated microglial synaptic phagocytosis, and astrocytic C3 activation. This intervention preserved dendritic spine density and ultimately reduced interictal spike frequency in EEG recordings in the chronic phase.

Our findings demonstrate that C1q acts as an upstream mediator of complement-driven neuronal loss during epileptogenesis. C1q blockade interrupted the pathogenic cascade involving microglial engulfment and astrocytic C3 upregulation, thereby preserving synaptic and neuronal integrity and reducing interictal spike activity. Our study highlights the therapeutic potential of C1q inhibition, which can provide multifaceted benefits by mitigating neuroinflammation, improving synaptic integrity, and reducing epileptiform activity during epileptogenesis.