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oai:docta.ucm.es:20.500.14352/1047982025-10-21T23:46:24Zcom_20.500.14352_14col_20.500.14352_15

00925njm 22002777a 4500 dc Alves, Mariana author Gil, Beatriz author Villegas Salmerón, Javier author Salari, Valentina author Martins Ferreira, Ricardo author Arribas Blázquez, Marina author Menéndez Méndez, Aida author Da Rosa Gerbatin, Rogerio author Smith, Jonathon author De Diego García, Laura author Conte, Giorgia author Sierra Márquez, Juan author Merino Serrais, Paula author Mitra, Meghma author Fernández Martín, Ana author Wang, Yitao author Kesavan, Jaideep author Melia, Ciara author Parras, Alberto author Beamer, Edward author Zimmer, Béla author Heiland, Mona author Cavanagh, Brenton author Parcianello Cipolat, Rafael author Morgan, James author Teng, Xinchen author Rodríguez Artalejo, Antonio author Olivos Ore, Luis Alcides author Engel, Tobias author 2024-05-20 Background: The purinergic ATP-gated P2X7 receptor (P2X7R) is increasingly recognized to contribute to pathological neuroinflammation and brain hyperexcitability. P2X7R expression has been shown to be increased in the brain, including both microglia and neurons, in experimental models of epilepsy and patients. To date, the cell type-specific downstream effects of P2X7Rs during seizures remain, however, incompletely understood. Methods: Effects of P2X7R signaling on seizures and epilepsy were analyzed in induced seizure models using male mice including the kainic acid model of status epilepticus and pentylenetetrazole model and in male and female mice in a genetic model of Dravet syndrome. RNA sequencing was used to analyze P2X7R downstream signaling during seizures. To investigate the cell type-specific role of the P2X7R during seizures and epilepsy, we generated mice lacking exon 2 of the P2rx7 gene in either microglia (P2rx7:Cx3cr1-Cre) or neurons (P2rx7:Thy-1-Cre). To investigate the protective potential of overexpressing P2X7R in GABAergic interneurons, P2X7Rs were overexpressed using adeno-associated virus transduction under the mDlx promoter. Results: RNA sequencing of hippocampal tissue from wild-type and P2X7R knock-out mice identified both glial and neuronal genes, in particular genes involved in GABAergic signaling, under the control of the P2X7R following seizures. Mice with deleted P2rx7 in microglia displayed less severe acute seizures and developed a milder form of epilepsy, and microglia displayed an anti-inflammatory molecular profile. In contrast, mice lacking P2rx7 in neurons showed a more severe seizure phenotype when compared to epileptic wild-type mice. Analysis of single-cell expression data revealed that human P2RX7 expression is elevated in the hippocampus of patients with temporal lobe epilepsy in excitatory and inhibitory neurons. Functional studies determined that GABAergic interneurons display increased responses to P2X7R activation in experimental epilepsy. Finally, we show that viral transduction of P2X7R in GABAergic interneurons protects against evoked and spontaneous seizures in experimental temporal lobe epilepsy and in mice lacking Scn1a, a model of Dravet syndrome. Conclusions: Our results suggest a dual and opposing action of P2X7R in epilepsy and suggest P2X7R overexpression in GABAergic interneurons as a novel therapeutic strategy for acquired and, possibly, genetic forms of epilepsy. Mariana Alves, Beatriz Gil, Javier Villegas-Salmerón, Valentina Salari, Ricardo Martins-Ferreira, Marina Arribas Blázquez, Aida Menéndez Méndez, Rogerio Da Rosa Gerbatin, Jonathon Smith, Laura de Diego-Garcia, Giorgia Conte, Juan Sierra-Marquez, Paula Merino Serrais, Meghma Mitra, Ana Fernandez Martin, Yitao Wang, Jaideep Kesavan, Ciara Melia, Alberto Parras, Edward Beamer, Béla Zimmer, Mona Heiland, Brenton Cavanagh, Rafael Parcianello Cipolat, James Morgan, Xinchen Teng, Jochen H.M. Prehn, Paolo F. Fabene, Giuseppe Bertini, Antonio R. Artalejo, Esteban Ballestar, Annette Nicke, Luis A. Olivos-Oré, Niamh M.C. Connolly, David C. Henshall, Tobias Engel, Opposing effects of the purinergic P2X7 receptor on seizures in neurons and microglia in male mice, Brain, Behavior, and Immunity, Volume 120, 2024, Pages 121-140, https://doi.org/10.1016/j.bbi.2024.05.023. 0889-1591 10.1016/j.bbi.2024.05.023 https://hdl.handle.net/20.500.14352/104798 1090-2139 https://doi.org/10.1016/j.bbi.2024.05.023 Opposing effects of the purinergic P2X7 receptor on seizures in neurons and microglia in male mice