Procedures for Culturing and Genetically Manipulating Murine Hippocampal Postnatal Neurons
Neuronal hippocampal cultures are simple and valuable models for studying neuronal function. While embryonic cultures are widely used for different applications, mouse postnatal cultures are still challenging, lack reproducibility and/or exhibit inappropriate neuronal activity. Yet, postnatal cultur...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-04-01
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| Series: | Frontiers in Synaptic Neuroscience |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fnsyn.2020.00019/full |
| _version_ | 1819296573621796864 |
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| author | Enora Moutin Anne-Laure Hemonnot Anne-Laure Hemonnot Vincent Seube Vincent Seube Nathalie Linck Nathalie Linck François Rassendren François Rassendren Julie Perroy Vincent Compan Vincent Compan |
| author_facet | Enora Moutin Anne-Laure Hemonnot Anne-Laure Hemonnot Vincent Seube Vincent Seube Nathalie Linck Nathalie Linck François Rassendren François Rassendren Julie Perroy Vincent Compan Vincent Compan |
| author_sort | Enora Moutin |
| collection | DOAJ |
| description | Neuronal hippocampal cultures are simple and valuable models for studying neuronal function. While embryonic cultures are widely used for different applications, mouse postnatal cultures are still challenging, lack reproducibility and/or exhibit inappropriate neuronal activity. Yet, postnatal cultures have major advantages such as allowing genotyping of pups before culture and reducing the number of experimental animals. Herein we describe a simple and fast protocol for culturing and genetically manipulating hippocampal neurons from P0 to P3 mice. This protocol provides reproducible cultures exhibiting a consistent neuronal development, normal excitatory over inhibitory neuronal ratio and a physiological neuronal activity. We also describe simple and efficient procedures for genetic manipulation of neurons using transfection reagent or lentiviral particles. Overall, this method provides a detailed and validated protocol allowing to explore cellular mechanisms and neuronal activity in postnatal hippocampal neurons in culture. |
| first_indexed | 2024-12-24T05:00:16Z |
| format | Article |
| id | doaj.art-44a820dedc644118aa4de2b29b7c24b1 |
| institution | Directory Open Access Journal |
| issn | 1663-3563 |
| language | English |
| last_indexed | 2024-12-24T05:00:16Z |
| publishDate | 2020-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Synaptic Neuroscience |
| spelling | doaj.art-44a820dedc644118aa4de2b29b7c24b12022-12-21T17:14:13ZengFrontiers Media S.A.Frontiers in Synaptic Neuroscience1663-35632020-04-011210.3389/fnsyn.2020.00019521872Procedures for Culturing and Genetically Manipulating Murine Hippocampal Postnatal NeuronsEnora Moutin0Anne-Laure Hemonnot1Anne-Laure Hemonnot2Vincent Seube3Vincent Seube4Nathalie Linck5Nathalie Linck6François Rassendren7François Rassendren8Julie Perroy9Vincent Compan10Vincent Compan11Institut de Génomique Fonctionnelle (IGF), University of Montpellier, CNRS, INSERM, Montpellier, FranceInstitut de Génomique Fonctionnelle (IGF), University of Montpellier, CNRS, INSERM, Montpellier, FranceLaboratoire d’Excellence Canaux Ioniques d’Intérêt Thérapeutique (LabEx ICST), Montpellier, FranceInstitut de Génomique Fonctionnelle (IGF), University of Montpellier, CNRS, INSERM, Montpellier, FranceLaboratoire d’Excellence Canaux Ioniques d’Intérêt Thérapeutique (LabEx ICST), Montpellier, FranceInstitut de Génomique Fonctionnelle (IGF), University of Montpellier, CNRS, INSERM, Montpellier, FranceLaboratoire d’Excellence Canaux Ioniques d’Intérêt Thérapeutique (LabEx ICST), Montpellier, FranceInstitut de Génomique Fonctionnelle (IGF), University of Montpellier, CNRS, INSERM, Montpellier, FranceLaboratoire d’Excellence Canaux Ioniques d’Intérêt Thérapeutique (LabEx ICST), Montpellier, FranceInstitut de Génomique Fonctionnelle (IGF), University of Montpellier, CNRS, INSERM, Montpellier, FranceInstitut de Génomique Fonctionnelle (IGF), University of Montpellier, CNRS, INSERM, Montpellier, FranceLaboratoire d’Excellence Canaux Ioniques d’Intérêt Thérapeutique (LabEx ICST), Montpellier, FranceNeuronal hippocampal cultures are simple and valuable models for studying neuronal function. While embryonic cultures are widely used for different applications, mouse postnatal cultures are still challenging, lack reproducibility and/or exhibit inappropriate neuronal activity. Yet, postnatal cultures have major advantages such as allowing genotyping of pups before culture and reducing the number of experimental animals. Herein we describe a simple and fast protocol for culturing and genetically manipulating hippocampal neurons from P0 to P3 mice. This protocol provides reproducible cultures exhibiting a consistent neuronal development, normal excitatory over inhibitory neuronal ratio and a physiological neuronal activity. We also describe simple and efficient procedures for genetic manipulation of neurons using transfection reagent or lentiviral particles. Overall, this method provides a detailed and validated protocol allowing to explore cellular mechanisms and neuronal activity in postnatal hippocampal neurons in culture.https://www.frontiersin.org/article/10.3389/fnsyn.2020.00019/fullneuronspostnatalhippocampusculturetransfectiontransduction |
| spellingShingle | Enora Moutin Anne-Laure Hemonnot Anne-Laure Hemonnot Vincent Seube Vincent Seube Nathalie Linck Nathalie Linck François Rassendren François Rassendren Julie Perroy Vincent Compan Vincent Compan Procedures for Culturing and Genetically Manipulating Murine Hippocampal Postnatal Neurons Frontiers in Synaptic Neuroscience neurons postnatal hippocampus culture transfection transduction |
| title | Procedures for Culturing and Genetically Manipulating Murine Hippocampal Postnatal Neurons |
| title_full | Procedures for Culturing and Genetically Manipulating Murine Hippocampal Postnatal Neurons |
| title_fullStr | Procedures for Culturing and Genetically Manipulating Murine Hippocampal Postnatal Neurons |
| title_full_unstemmed | Procedures for Culturing and Genetically Manipulating Murine Hippocampal Postnatal Neurons |
| title_short | Procedures for Culturing and Genetically Manipulating Murine Hippocampal Postnatal Neurons |
| title_sort | procedures for culturing and genetically manipulating murine hippocampal postnatal neurons |
| topic | neurons postnatal hippocampus culture transfection transduction |
| url | https://www.frontiersin.org/article/10.3389/fnsyn.2020.00019/full |
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