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Monosynaptic tracing: a step-by-step protocol.

Thomas K Lavin1, Lei Jin1, Ian R Wickersham1

  • 1McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA, United States.

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|August 14, 2019
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Summary
This summary is machine-generated.

This study details a method for mapping neural connections using rabies virus tracing. It provides a step-by-step protocol for whole-brain analysis of presynaptic neurons, crucial for understanding neural circuits.

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Genetics

Background:

  • Understanding neural circuitry requires mapping neuronal connections.
  • Monosynaptic tracing techniques are essential for identifying direct neuronal inputs.
  • Rabies virus-based tracing offers a powerful tool for this purpose.

Purpose of the Study:

  • To present a detailed protocol for monosynaptic tracing using deletion-mutant rabies virus.
  • To enable whole-brain mapping of neurons directly presynaptic to a targeted population.
  • To provide a robust method leveraging Cre mouse lines and adeno-associated viral vectors.

Main Methods:

  • Utilizing Cre-dependent adeno-associated viral vectors for gene expression in targeted neurons.
  • Employing first-generation (ΔG) rabies viral vectors for tracing.
  • Performing step-by-step experimental procedures for rabies virus tracing.

Main Results:

  • The protocol facilitates the identification of monosynaptic inputs to specific neuronal populations.
  • Whole-brain mapping of presynaptic partners is achievable with this method.
  • The described approach is robust and reproducible.

Conclusions:

  • This protocol provides a standardized method for monosynaptic rabies virus tracing.
  • It enables detailed mapping of neural connectivity within the brain.
  • The technique is valuable for advancing the study of neural circuits.