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ESTABLISHMENT OF AN OPTIMIZED AND AUTOMATED WORKFLOW FOR WHOLE BRAIN PROBING OF NEURONAL ACTIVITY PATTERNS IN TRAP MICE
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Edité par CCSD -
International audience. Aims: Behaviors are encoded by neural circuits that are widespread within the brain and change with age and experience. Immunodetection of the immediate early gene c-Fos has been successfully used for decades to reveal neural circuits active during specific tasks or conditions. Our aims here is to develop a workflow that circumvents classical temporal and spatial limitations associated to c-Fos quantifications.Method: We used a new genetic TRAP method combining Cre-dependent tdTomato expression under c-Fos promoter with c-Fos immunohistochemistry, allowing visualization and direct comparison of neural circuits activated at different times or during different tasks. We established a workflow that optimize and automate cell detection, cell classification (e.g. c-Fos vs. c-Fos/tdTomato) and whole brain registration.Results: We demonstrate that this workflow allows accurate cell number quantification with minimal interindividual variability. Further, interrogation of brain atlases at different scales (from simplified to detailed) was achieved allowing gradually zooming on brain regions to explore spatial distribution of activated cells. Finally, we illustrate the potential of this approach by comparing patterns of neuronal activation in various contexts (e. g. different vigilance states, complex behavioral tasks …).Conclusion: Altogether, this automated workflow allows a fast and accurate analysis of whole brain activity pattern at the cellular level, in various contexts.
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