title: High-Throughput LC-MS/MS Proteomic Analysis of a Mouse Model of Mesiotemporal Lobe Epilepsy Predicts Microglial Activation Underlying Disease Development.
abstract: Uncovering the molecular mechanisms of mesiotemporal lobe epilepsy (MTLE) is critical to identify therapeutic targets. In this study, we performed global protein expression analysis of a kainic acid (KA) MTLE mouse model at various time-points (1, 3, and 30 days post-KA injection -dpi), representing specific stages of the syndrome. High-resolution liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), in combination with label-free protein quantification using three processing approaches for quantification, was applied. Following comparison of KA versus NaCl-injected mice, 22, 53, and 175 proteins were differentially (statistically significant) expressed at 1, 3 and 30dpi, respectively, according to all three quantification approaches. Selected findings were confirmed by multiple reaction monitoring LC-MS/MS. As a positive control, the astrocyte marker GFAP was found to be upregulated (3dpi: 1.9 fold; 30dpi: 12.5 fold), also verified by IHC. The results collectively suggest that impairment in synaptic transmission occurs even right after initial status epilepticus (1dpi), with neurodegeneration becoming more extensive during epileptogenesis (3dpi) and sustained at the chronic phase (30dpi), where also extensive glial- and astrocyte-mediated inflammation is evident. This molecular profile is in line with observed phenotypic changes in human MTLE, providing the basis for future studies on new molecular targets for the disease.
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens , Soranou Efessiou 4, 11527 Athens, Greece.
authors.forename: Vasiliki
authors.initials: V
authors.lastname: Bitsika
authors.affiliation: SynapCell SAS , F-38700 La Tronche, France.
authors.forename: Venceslas
authors.initials: V
authors.lastname: Duveau
authors.affiliation: Inserm, U1216, Grenoble-Institut des Neurosciences , F-38000 Grenoble, France.
authors.forename: Julia
authors.initials: J
authors.lastname: Simon-Areces
authors.affiliation: BHF Glasgow Cardiovascular Research Centre, University of Glasgow , G12 8QQ Glasgow, United Kingdom.
authors.forename: William
authors.initials: W
authors.lastname: Mullen
authors.affiliation: SynapCell SAS , F-38700 La Tronche, France.
authors.forename: Corinne
authors.initials: C
authors.lastname: Roucard
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens , Soranou Efessiou 4, 11527 Athens, Greece.
authors.forename: Manousos
authors.initials: M
authors.lastname: Makridakis
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens , Soranou Efessiou 4, 11527 Athens, Greece.
authors.forename: George
authors.initials: G
authors.lastname: Mermelekas
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens , Soranou Efessiou 4, 11527 Athens, Greece.
authors.forename: Pantelis
authors.initials: P
authors.lastname: Savvopoulos
authors.affiliation: Inserm, U1216, Grenoble-Institut des Neurosciences , F-38000 Grenoble, France.
authors.forename: Antoine
authors.initials: A
authors.lastname: Depaulis
authors.affiliation: Biotechnology Division, Biomedical Research Foundation, Academy of Athens , Soranou Efessiou 4, 11527 Athens, Greece.
authors.forename: Antonia
authors.initials: A
authors.lastname: Vlahou
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D004195:Disease Models, Animal; D018450:Disease Progression; D004833:Epilepsy, Temporal Lobe / Q000209:etiology / Q000473:pathology* / Q000503:physiopathology; D057166:High-Throughput Screening Assays; D007608:Kainic Acid; D051379:Mice; D017628:Microglia / Q000378:metabolism*; D019636:Neurodegenerative Diseases; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D009435:Synaptic Transmission; D053719:Tandem Mass Spectrometry; D013997:Time Factors
chemical_list: D020543:Proteome; D007608:Kainic Acid
keywords: LC−MS/MS; MTLE; epilepsy; kainic acid; proteomics