Diisopropylfluorophosphate-induced status epilepticus drives complex glial cell phenotypes in adult male mice

Organophosphate pesticides and nerve agents (OPs), are characterized by cholinesterase inhibition. In addition to severe peripheral symptoms, high doses of OPs can lead to seizures and status epilepticus (SE). Long lasting seizure activity and subsequent neurodegeneration promote neuroinflammation l...

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Main Authors: Clémence Maupu, Julie Enderlin, Alexandre Igert, Myriam Oger, Stéphane Auvin, Rahma Hassan-Abdi, Nadia Soussi-Yanicostas, Xavier Brazzolotto, Florian Nachon, Grégory Dal Bo, Nina Dupuis
Format: Article
Language:English
Published: Elsevier 2021-05-01
Series:Neurobiology of Disease
Subjects:
DFP
Online Access:http://www.sciencedirect.com/science/article/pii/S0969996121000255
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language English
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author Clémence Maupu
Julie Enderlin
Alexandre Igert
Myriam Oger
Stéphane Auvin
Rahma Hassan-Abdi
Nadia Soussi-Yanicostas
Xavier Brazzolotto
Florian Nachon
Grégory Dal Bo
Nina Dupuis
spellingShingle Clémence Maupu
Julie Enderlin
Alexandre Igert
Myriam Oger
Stéphane Auvin
Rahma Hassan-Abdi
Nadia Soussi-Yanicostas
Xavier Brazzolotto
Florian Nachon
Grégory Dal Bo
Nina Dupuis
Diisopropylfluorophosphate-induced status epilepticus drives complex glial cell phenotypes in adult male mice
Neurobiology of Disease
Astrocyte
Microglia
Cytokines
Seizure
DFP
Organophosphates
author_facet Clémence Maupu
Julie Enderlin
Alexandre Igert
Myriam Oger
Stéphane Auvin
Rahma Hassan-Abdi
Nadia Soussi-Yanicostas
Xavier Brazzolotto
Florian Nachon
Grégory Dal Bo
Nina Dupuis
author_sort Clémence Maupu
title Diisopropylfluorophosphate-induced status epilepticus drives complex glial cell phenotypes in adult male mice
title_short Diisopropylfluorophosphate-induced status epilepticus drives complex glial cell phenotypes in adult male mice
title_full Diisopropylfluorophosphate-induced status epilepticus drives complex glial cell phenotypes in adult male mice
title_fullStr Diisopropylfluorophosphate-induced status epilepticus drives complex glial cell phenotypes in adult male mice
title_full_unstemmed Diisopropylfluorophosphate-induced status epilepticus drives complex glial cell phenotypes in adult male mice
title_sort diisopropylfluorophosphate-induced status epilepticus drives complex glial cell phenotypes in adult male mice
publisher Elsevier
series Neurobiology of Disease
issn 1095-953X
publishDate 2021-05-01
description Organophosphate pesticides and nerve agents (OPs), are characterized by cholinesterase inhibition. In addition to severe peripheral symptoms, high doses of OPs can lead to seizures and status epilepticus (SE). Long lasting seizure activity and subsequent neurodegeneration promote neuroinflammation leading to profound pathological alterations of the brain.The aim of this study was to characterize neuroinflammatory responses at key time points after SE induced by the OP, diisopropylfluorophosphate (DFP). Immunohistochemistry (IHC) analysis and RT-qPCR on cerebral tissue are often insufficient to identity and quantify precise neuroinflammatory alterations. To address these needs, we performed RT-qPCR quantification after whole brain magnetic-activated cell-sorting (MACS) of CD11B (microglia/infiltrated macrophages) and GLAST (astrocytes)-positive cells at 1, 4, 24 h and 3 days post-SE. In order to compare these results to those obtained by IHC, we performed, classical Iba1 (microglia/infiltrated macrophages) and GFAP (astrocytes) IHC analysis in parallel, focusing on the hippocampus, a brain region affected by seizure activity and neurodegeneration.Shortly after SE (1–4 h), an increase in pro-inflammatory (M1-like) markers and A2-specific markers, proposed as neurotrophic, were observed in CD11B and GLAST-positive isolated cells, respectively. Microglial cells successively expressed immuno-regulatory (M2b-like) and anti-inflammatory (M2a-like) at 4 h and 24 h post-SE induction. At 24 h and 3 days, A1-specific markers, proposed as neurotoxic, were increased in isolated astrocytes. Although IHC analysis presented no modification in terms of percentage of marked area and cell number at 1 and 4 h after SE, at 24 h and 3 days after SE, microglial and astrocytic activation was visible by IHC as an increase in Iba1 and GFAP-positive area and Iba1-positive cells in DFP animals when compared to the control.Our work identified sequential microglial and astrocytic phenotype activation. Although the role of each phenotype in SE cerebral outcomes requires further study, targeting specific markers at specific time point could be a beneficial strategy for DFP-induced SE treatment.
topic Astrocyte
Microglia
Cytokines
Seizure
DFP
Organophosphates
url http://www.sciencedirect.com/science/article/pii/S0969996121000255
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spelling doaj-a2925ef027ff413ab4ff53a35214c3ee2021-03-22T08:43:09ZengElsevierNeurobiology of Disease1095-953X2021-05-01152105276Diisopropylfluorophosphate-induced status epilepticus drives complex glial cell phenotypes in adult male miceClémence Maupu0Julie Enderlin1Alexandre Igert2Myriam Oger3Stéphane Auvin4Rahma Hassan-Abdi5Nadia Soussi-Yanicostas6Xavier Brazzolotto7Florian Nachon8Grégory Dal Bo9Nina Dupuis10Département de Toxicologie et risques chimiques, Institut de recherche biomédicale des armées, BP73, F-91223 Brétigny sur Orge cedex, FranceUniversité de Paris, NeuroDiderot, Inserm, F-75019 Paris, France; Service de neurologie pédiatrique, AP-HP, Hôpital Robert Debré, F-75019 Paris, FranceDépartement de Toxicologie et risques chimiques, Institut de recherche biomédicale des armées, BP73, F-91223 Brétigny sur Orge cedex, FranceUnité Imagerie, Institut de recherche biomédicale des armées, BP73, F-91223 Brétigny sur Orge cedex, FranceUniversité de Paris, NeuroDiderot, Inserm, F-75019 Paris, France; Service de neurologie pédiatrique, AP-HP, Hôpital Robert Debré, F-75019 Paris, FranceUniversité de Paris, NeuroDiderot, Inserm, F-75019 Paris, FranceUniversité de Paris, NeuroDiderot, Inserm, F-75019 Paris, FranceDépartement de Toxicologie et risques chimiques, Institut de recherche biomédicale des armées, BP73, F-91223 Brétigny sur Orge cedex, FranceDépartement de Toxicologie et risques chimiques, Institut de recherche biomédicale des armées, BP73, F-91223 Brétigny sur Orge cedex, FranceDépartement de Toxicologie et risques chimiques, Institut de recherche biomédicale des armées, BP73, F-91223 Brétigny sur Orge cedex, FranceDépartement de Toxicologie et risques chimiques, Institut de recherche biomédicale des armées, BP73, F-91223 Brétigny sur Orge cedex, France; Corresponding author at: Institut de recherche biomédicale des armées, 1 Place Général Valérie André, BP 73, 91223 Brétigny sur Orge cedex, France.Organophosphate pesticides and nerve agents (OPs), are characterized by cholinesterase inhibition. In addition to severe peripheral symptoms, high doses of OPs can lead to seizures and status epilepticus (SE). Long lasting seizure activity and subsequent neurodegeneration promote neuroinflammation leading to profound pathological alterations of the brain.The aim of this study was to characterize neuroinflammatory responses at key time points after SE induced by the OP, diisopropylfluorophosphate (DFP). Immunohistochemistry (IHC) analysis and RT-qPCR on cerebral tissue are often insufficient to identity and quantify precise neuroinflammatory alterations. To address these needs, we performed RT-qPCR quantification after whole brain magnetic-activated cell-sorting (MACS) of CD11B (microglia/infiltrated macrophages) and GLAST (astrocytes)-positive cells at 1, 4, 24 h and 3 days post-SE. In order to compare these results to those obtained by IHC, we performed, classical Iba1 (microglia/infiltrated macrophages) and GFAP (astrocytes) IHC analysis in parallel, focusing on the hippocampus, a brain region affected by seizure activity and neurodegeneration.Shortly after SE (1–4 h), an increase in pro-inflammatory (M1-like) markers and A2-specific markers, proposed as neurotrophic, were observed in CD11B and GLAST-positive isolated cells, respectively. Microglial cells successively expressed immuno-regulatory (M2b-like) and anti-inflammatory (M2a-like) at 4 h and 24 h post-SE induction. At 24 h and 3 days, A1-specific markers, proposed as neurotoxic, were increased in isolated astrocytes. Although IHC analysis presented no modification in terms of percentage of marked area and cell number at 1 and 4 h after SE, at 24 h and 3 days after SE, microglial and astrocytic activation was visible by IHC as an increase in Iba1 and GFAP-positive area and Iba1-positive cells in DFP animals when compared to the control.Our work identified sequential microglial and astrocytic phenotype activation. Although the role of each phenotype in SE cerebral outcomes requires further study, targeting specific markers at specific time point could be a beneficial strategy for DFP-induced SE treatment.http://www.sciencedirect.com/science/article/pii/S0969996121000255AstrocyteMicrogliaCytokinesSeizureDFPOrganophosphates