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Protocol to visualize three distinct neuronal ensembles encoding different events in the mouse brain using genetic and viral approaches

  • 0Program in Neurosciences & Mental Health, Hospital for Sick Children, 555 University Avenue, Toronto, ON M5G 1X8, Canada; Department of Cellular and Computational Neuroscience, Swammerdam Institute for Life Sciences, Amsterdam Neuroscience, University of Amsterdam, Amsterdam 1090 GE, the Netherlands.
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April 9, 2025

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April 9, 2025

View abstract on PubMed

Summary

This summary is machine-generated.

This study presents a new protocol for visualizing active neuronal ensembles during memory phases in mice. The method uses viral vectors and fluorescent proteins for detailed analysis of neural activity and memory events.

Area Of Science

  • Neuroscience
  • Molecular Biology
  • Genetics

Background

  • Activity tagging is crucial for understanding neuronal function.
  • Visualizing distinct neuronal ensembles is key to studying memory formation.
  • Existing methods may lack the precision for multi-phase memory analysis.

Purpose Of The Study

  • To provide a detailed protocol for visualizing separate neuronal ensembles active during distinct memory phases.
  • To enable the study of neural circuit dynamics across different stages of memory.
  • To offer a robust method for activity-dependent neuronal labeling in transgenic mice.

Main Methods

  • Utilizing viral microinjection to deliver genetic constructs.
  • Employing a robust activity marker (RAM) promoter for GFP expression in active neurons.
  • Using transgenic mice expressing tdTomato (TdT) for co-labeling.
  • Performing immunohistochemical (IHC) visualization of endogenous cFos expression.
  • Detailing tissue preparation, imaging, and quantification procedures.

Main Results

  • Successful visualization of distinct neuronal ensembles associated with specific memory phases.
  • Demonstration of co-labeling strategies for enhanced specificity.
  • Establishment of a comprehensive workflow from viral delivery to data analysis.
  • Quantification of memory-related neural activity patterns.

Conclusions

  • The presented protocol offers a powerful tool for dissecting neural ensembles involved in memory.
  • This method facilitates the detailed study of neural circuit activity during complex cognitive processes.
  • The protocol is adaptable for various research questions in systems neuroscience and memory research.

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