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JoVE Journal
Neuroscience
Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenet...
Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenet...
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex

Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex

Full Text
20,892 Views
10:45 min
December 19, 2012

DOI: 10.3791/4328-v

William C. Clegern1, Michele E. Moore1, Michelle A. Schmidt1, Jonathan Wisor1

1Department of Veterinary & Comparative Anatomy, Pharmacology and Physiology, Sleep and Performance Research Center, WWAMI Medical Education Program,Washington State University

Overview

This procedure outlines the manipulation of cerebral cortical pyramidal neurons using optogenetics while monitoring electroencephalogram (EEG), electromyogram (EMG), and cerebral lactate concentration. The experimental setup is conducted on cable-tethered mice during spontaneous sleep/wake cycles.

Key Study Components

Area of Science

  • Neuroscience
  • Optogenetics
  • Electrophysiology

Background

  • Optogenetics allows precise control of neuronal activity.
  • Monitoring brain biochemistry and electrophysiology in real-time is crucial.
  • EEG and lactate concentration provide insights into neuronal function.
  • This method offers advantages over traditional pharmacological studies.

Purpose of Study

  • To measure EEG-defined sleep and lactate concentration during optogenetic stimulation.
  • To manipulate neuronal activity at precise stereotaxic coordinates.
  • To collect data on brain activity while the animal behaves naturally.

Methods Used

  • Surgically implant electroencephalographic leads and cannulas for lactate sensors.
  • Insert fiber optic cable and pre-calibrated lactate sensor into guide cannulas.
  • Adjust blue light stimulus intensity for desired electrophysiological response.
  • Collect EEG, EMG, and lactate concentration data during spontaneous behavior.

Main Results

  • Real-time monitoring of brain biochemistry and electrophysiology achieved.
  • Successful manipulation of neuronal activity with optogenetic stimulation.
  • Data collected during natural sleep/wake cycles.
  • Enhanced understanding of neuronal function and metabolic processes.

Conclusions

  • This method provides a powerful tool for studying neuronal dynamics.
  • Real-time data collection enhances insights into brain function.
  • Optogenetic stimulation combined with biochemical monitoring is effective.

Frequently Asked Questions

What is optogenetics?
Optogenetics is a technique that uses light to control neurons that have been genetically modified to express light-sensitive ion channels.
How does this method differ from pharmacological studies?
Unlike pharmacological studies, this method allows for real-time manipulation of neuronal activity and monitoring of brain biochemistry simultaneously.
What are the advantages of using EEG in this study?
EEG provides a non-invasive way to monitor electrical activity in the brain, allowing researchers to correlate neuronal activity with behavioral states.
What is the significance of measuring lactate concentration?
Lactate concentration can indicate metabolic activity in the brain and is important for understanding energy metabolism during neuronal activity.
What types of data are collected during the experiment?
Data collected includes electroencephalographic (EEG) signals, electromyographic (EMG) signals, and cerebral lactate concentration.
Can this method be applied to other types of neurons?
Yes, the method can potentially be adapted to study other types of neurons by modifying the optogenetic tools used.

A procedure is described for manipulating the activity of cerebral cortical pyramidal neurons optogenetically while the electroencephalogram, electromyogram, and cerebral lactate concentration are monitored. Experimental recordings are performed on cable-tethered mice while they undergo spontaneous sleep/wake cycles. Optogenetic equipment is assembled in our laboratory; recording equipment is commercially available.

The overall aim of this procedure is to measure EEG defined sleep and lactate concentration during optogenetic stimulation at precise stereotaxic coordinates. Surgically implant the necessary electroencephalographic leads and cannulas for a lactate sensor and fiber optic cable. Then insert the fiber optic cable and pre calibrated lactate sensor into their guide cannulas in the animal's skull.

Next, adjust the blue light stimulus intensity to achieve the desired electrophysiological response. Then proceed to collect the electroencephalographic, electromyographic and lactate concentration data while the animal is allowed to spontaneously behave and or sleep. Compared to other methods like pharmacological studies of the EEG, this experimental approach has the advantage of monitoring brain biochemistry and electrophysiology while the activity of neurons is manipulated in real time on a sub-second time scale.

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