Temporal Dynamics of Neocortical Development in Organotypic Mouse Cultures: A Comprehensive Analysis

Author:

Bak AniellaORCID,Schmied KatharinaORCID,Jakob MortenORCID,Bedogni FrancescoORCID,Squire Olivia,Gittel Birgit,Jesinghausen MaikORCID,Schünemann KerstinORCID,Weber YvonneORCID,Kampa BjörnORCID,van Loo Karen M. J.ORCID,Koch HennerORCID

Abstract

AbstractMurine organotypic brain slice cultures have been widely used in neuroscientific research and are offering the opportunity to study neuronal function under normal and disease conditions. Despite the brought application, the mechanisms governing the maturation of immature cortical circuitsin vitroare not well understood. In this study, we present a detailed investigation into the development of the neocortexin vitro. Utilizing a holistic approach, we studied organotypic whole-hemisphere brain slice cultures from postnatal mice and tracked the development of the somatosensory area over a five-week period. Our analysis revealed the maturation of passive and active intrinsic properties of pyramidal cells together with their morphology, closely resemblingin vivodevelopment. Detailed Multi-electrode array (MEA) electrophysiological assessments and RNA expression profiling demonstrated stable network properties by two weeks in culture, followed by the transition of spontaneous activity towards more complex patterns including high-frequency oscillations. However, weeks 4 and 5 exhibited increased variability and initial signs of neuronal loss, highlighting the importance of considering developmental stages in experimental design. This comprehensive characterization is vital for understanding the temporal dynamics of the neocortical developmentin vitro, with implications for neuroscientific research methodologies, particularly in the investigation of diseases such as epilepsy and other neurodevelopmental disorders.

Publisher

Cold Spring Harbor Laboratory

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