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Combining Optogenetics with Artificial microRNAs to Characterize the Effects of Gene Knockdown on Presynaptic Function within Intact Neuronal Circuits
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Combining Optogenetics with Artificial microRNAs to Characterize the Effects of Gene Knockdown on Presynaptic Function within Intact Neuronal Circuits

Combining Optogenetics with Artificial microRNAs to Characterize the Effects of Gene Knockdown on Presynaptic Function within Intact Neuronal Circuits

DOI:
10.3791/57223-v

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09:17 min

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March 14, 2018

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, ,
1Center for Synaptic Neuroscience and Technology,Istituto Italiano di Tecnologia

Chapters

  • 00:04Title
  • 01:09Extracting RNA from Primary Cultures to Evaluate microRNA Knockdown Efficiency
  • 03:53Stimulation of Knocked-down Neurons Using Optogenetics
  • 07:13Results: Knockdown of P/Q-type Voltage-gated Ca2+ Channel at CA3 to CA1 Synapses
  • 08:36Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol provides a workflow on how to combine artificial microRNA-mediated RNA interference with optogenetics to stimulate specifically presynaptic boutons with reduced expression of selective gene(s) within intact neuronal circuits.

Tags

Keywords: OptogeneticsArtificial MicroRNARNA InterferencePresynaptic FunctionNeuronal CircuitsSynaptic PhysiologyPresynaptic ProteinsNeuronal CultureGene KnockdownAcute Brain SlicesIn VivoCorticoculturingAraCRNA ExtractionChloroform

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