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Optogenetic Methods for the Study of Circadian Rhythms.

Jeff R Jones1,2, Michael C Tackenberg1, Douglas G McMahon3,4

  • 1Vanderbilt Brain Institute, Vanderbilt University, Nashville, TN, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 7, 2020
PubMed
Summary

Optogenetics precisely manipulates electrical activity in mammalian clock neurons, revealing insights into circadian rhythms and behavior. This technique aids in understanding the timing signals generated by these crucial biological clocks.

Keywords:
BehaviorBioluminescenceChannelrhodopsinCircadianFiring rateIn vivoOptogenetics

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Area of Science:

  • Neuroscience
  • Chronobiology
  • Molecular Biology

Background:

  • Circadian clock neurons exhibit rhythmic spontaneous spike frequencies crucial for timing and rhythm generation.
  • Understanding the role of neuronal electrical activity in circadian timing is essential.
  • Optogenetics offers a powerful tool to investigate clock neuron function.

Purpose of the Study:

  • To describe protocols for optogenetic manipulation of mammalian clock neurons in the suprachiasmatic nucleus (SCN).
  • To investigate the roles of SCN clock neuron electrical activity in circadian physiology and behavior.
  • To enable precise control over neuronal firing rates within the SCN.

Main Methods:

  • Utilized in vitro and in vivo optogenetics targeting mammalian SCN clock neurons.
  • Employed channelrhodopsin for optogenetic stimulation and halorhodopsin for inhibition.
  • Combined optogenetic manipulation with actigraphy and gene expression analysis.

Main Results:

  • Demonstrated precise manipulation of neuronal firing rates in SCN clock neurons.
  • Enabled targeted control within specific neuronal subpopulations of the SCN.
  • Facilitated the study of circadian physiology and behavior through optogenetic intervention.

Conclusions:

  • Optogenetics provides a robust method for dissecting the function of SCN clock neuron electrical activity.
  • These protocols allow for detailed investigation into the mechanisms of circadian rhythm generation and output.
  • The combined approach offers new avenues for understanding circadian disorders and developing therapeutic strategies.