AUTHOR=Moutin Enora , Hemonnot Anne-Laure , Seube Vincent , Linck Nathalie , Rassendren François , Perroy Julie , Compan Vincent TITLE=Procedures for Culturing and Genetically Manipulating Murine Hippocampal Postnatal Neurons JOURNAL=Frontiers in Synaptic Neuroscience VOLUME=12 YEAR=2020 URL=https://www.frontiersin.org/journals/synaptic-neuroscience/articles/10.3389/fnsyn.2020.00019 DOI=10.3389/fnsyn.2020.00019 ISSN=1663-3563 ABSTRACT=

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.