Ex vivo model of epilepsy in organotypic slices—a new tool for drug screening
Abstract Background Epilepsy is a prevalent neurological disorder worldwide. It is characterized by an enduring predisposition to generate seizures and its development is accompanied by alterations in many cellular processes. Organotypic slice cultures represent a multicellular environment with the...
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| Format: | Article |
| Language: | English |
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BMC
2018-07-01
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| Series: | Journal of Neuroinflammation |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12974-018-1225-2 |
| _version_ | 1818031769142689792 |
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| author | Daniela M. Magalhães Noémia Pereira Diogo M. Rombo Cláudia Beltrão-Cavacas Ana M. Sebastião Cláudia A. Valente |
| author_facet | Daniela M. Magalhães Noémia Pereira Diogo M. Rombo Cláudia Beltrão-Cavacas Ana M. Sebastião Cláudia A. Valente |
| author_sort | Daniela M. Magalhães |
| collection | DOAJ |
| description | Abstract Background Epilepsy is a prevalent neurological disorder worldwide. It is characterized by an enduring predisposition to generate seizures and its development is accompanied by alterations in many cellular processes. Organotypic slice cultures represent a multicellular environment with the potential to assess biological mechanisms, and they are used as a starting point for refining molecules for in vivo studies. Here, we investigated organotypic slice cultures as a model of epilepsy. Methods We assessed, by electrophysiological recordings, the spontaneous activity of organotypic slices maintained under different culture protocols. Moreover, we evaluated, through molecular-based approaches, neurogenesis, neuronal death, gliosis, expression of proinflammatory cytokines, and activation of NLRP3 inflammasome (nucleotide-binding, leucine-rich repeat, pyrin domain) as biomarkers of neuroinflammation. Results We demonstrated that organotypic slices, maintained under a serum deprivation culture protocol, develop epileptic-like activity. Furthermore, throughout a comparative study with slices that do not depict any epileptiform activity, slices with epileptiform activity were found to display significant differences in terms of inflammation-related features, such as (1) increased neuronal death, with higher incidence in CA1 pyramidal neurons of the hippocampus; (2) activation of astrocytes and microglia, assessed through western blot and immunohistochemistry; (3) upregulation of proinflammatory cytokines, specifically interleukin-1β (IL-1β), interleukin-6, and tumor necrosis factor α, revealed by qPCR; and (4) enhanced expression of NLRP3, assessed by western blot, together with increased NLRP3 activation, showed by IL-1β quantification. Conclusions Thus, organotypic slice cultures gradually deprived of serum mimic the epileptic-like activity, as well as the inflammatory events associated with in vivo epilepsy. This system can be considered a new tool to explore the interplay between neuroinflammation and epilepsy and to screen potential drug candidates, within the inflammatory cascades, to reduce/halt epileptogenesis. |
| first_indexed | 2024-12-10T05:56:44Z |
| format | Article |
| id | doaj.art-55ac221e3d5a4e73b6e40ca5627291a0 |
| institution | Directory Open Access Journal |
| issn | 1742-2094 |
| language | English |
| last_indexed | 2024-12-10T05:56:44Z |
| publishDate | 2018-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Neuroinflammation |
| spelling | doaj.art-55ac221e3d5a4e73b6e40ca5627291a02022-12-22T01:59:54ZengBMCJournal of Neuroinflammation1742-20942018-07-0115111810.1186/s12974-018-1225-2Ex vivo model of epilepsy in organotypic slices—a new tool for drug screeningDaniela M. Magalhães0Noémia Pereira1Diogo M. Rombo2Cláudia Beltrão-Cavacas3Ana M. Sebastião4Cláudia A. Valente5Instituto de Farmacologia e Neurociências, Faculdade de Medicina, Universidade de LisboaInstituto de Farmacologia e Neurociências, Faculdade de Medicina, Universidade de LisboaInstituto de Farmacologia e Neurociências, Faculdade de Medicina, Universidade de LisboaInstituto de Farmacologia e Neurociências, Faculdade de Medicina, Universidade de LisboaInstituto de Farmacologia e Neurociências, Faculdade de Medicina, Universidade de LisboaInstituto de Farmacologia e Neurociências, Faculdade de Medicina, Universidade de LisboaAbstract Background Epilepsy is a prevalent neurological disorder worldwide. It is characterized by an enduring predisposition to generate seizures and its development is accompanied by alterations in many cellular processes. Organotypic slice cultures represent a multicellular environment with the potential to assess biological mechanisms, and they are used as a starting point for refining molecules for in vivo studies. Here, we investigated organotypic slice cultures as a model of epilepsy. Methods We assessed, by electrophysiological recordings, the spontaneous activity of organotypic slices maintained under different culture protocols. Moreover, we evaluated, through molecular-based approaches, neurogenesis, neuronal death, gliosis, expression of proinflammatory cytokines, and activation of NLRP3 inflammasome (nucleotide-binding, leucine-rich repeat, pyrin domain) as biomarkers of neuroinflammation. Results We demonstrated that organotypic slices, maintained under a serum deprivation culture protocol, develop epileptic-like activity. Furthermore, throughout a comparative study with slices that do not depict any epileptiform activity, slices with epileptiform activity were found to display significant differences in terms of inflammation-related features, such as (1) increased neuronal death, with higher incidence in CA1 pyramidal neurons of the hippocampus; (2) activation of astrocytes and microglia, assessed through western blot and immunohistochemistry; (3) upregulation of proinflammatory cytokines, specifically interleukin-1β (IL-1β), interleukin-6, and tumor necrosis factor α, revealed by qPCR; and (4) enhanced expression of NLRP3, assessed by western blot, together with increased NLRP3 activation, showed by IL-1β quantification. Conclusions Thus, organotypic slice cultures gradually deprived of serum mimic the epileptic-like activity, as well as the inflammatory events associated with in vivo epilepsy. This system can be considered a new tool to explore the interplay between neuroinflammation and epilepsy and to screen potential drug candidates, within the inflammatory cascades, to reduce/halt epileptogenesis.http://link.springer.com/article/10.1186/s12974-018-1225-2EpilepsyOrganotypic slice culturesNeuroinflammationGliosisProinflammatory cytokinesInterleukin-1β |
| spellingShingle | Daniela M. Magalhães Noémia Pereira Diogo M. Rombo Cláudia Beltrão-Cavacas Ana M. Sebastião Cláudia A. Valente Ex vivo model of epilepsy in organotypic slices—a new tool for drug screening Journal of Neuroinflammation Epilepsy Organotypic slice cultures Neuroinflammation Gliosis Proinflammatory cytokines Interleukin-1β |
| title | Ex vivo model of epilepsy in organotypic slices—a new tool for drug screening |
| title_full | Ex vivo model of epilepsy in organotypic slices—a new tool for drug screening |
| title_fullStr | Ex vivo model of epilepsy in organotypic slices—a new tool for drug screening |
| title_full_unstemmed | Ex vivo model of epilepsy in organotypic slices—a new tool for drug screening |
| title_short | Ex vivo model of epilepsy in organotypic slices—a new tool for drug screening |
| title_sort | ex vivo model of epilepsy in organotypic slices a new tool for drug screening |
| topic | Epilepsy Organotypic slice cultures Neuroinflammation Gliosis Proinflammatory cytokines Interleukin-1β |
| url | http://link.springer.com/article/10.1186/s12974-018-1225-2 |
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