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Neuroscience
Optogenetics Identification d’un Type Neuronal avec un verre de Optrode chez les souris éveillés
Optogenetics Identification d’un Type Neuronal avec un verre de Optrode chez les souris éveillés
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Optogenetics Identification of a Neuronal Type with a Glass Optrode in Awake Mice

Optogenetics Identification d’un Type Neuronal avec un verre de Optrode chez les souris éveillés

Full Text
7,244 Views
07:51 min
June 28, 2018

DOI: 10.3791/57781-v

Munenori Ono1, Shinji Muramoto1, Lanlan Ma1, Nobuo Kato1

1Department of Physiology,Kanazawa Medical University

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This study presents a method for optogenetic single-unit recording in awake mice using a custom glass optrode. The technique aims to provide insights into the functions of various neuron types by enabling reliable in vivo identification and manipulation with cost-effective materials.

Key Study Components

Area of Science

  • Neuroscience
  • Electrophysiology
  • Optogenetics

Background

  • The method focuses on understanding neuron functions, particularly GABAergic and glutamatergic neurons.
  • It utilizes a transgenic mouse model to express channelrhodopsin-II in inhibitory neurons.
  • Previous methods lacked reliability in in vivo neuron type identification.
  • Cost-effective and accessible materials are used in the methodology.

Purpose of Study

  • To develop a reliable technique for single-unit neuronal recordings in awake mice.
  • To investigate the functional roles of different neuron types during recorded activities.
  • To advance the tools available for studying neural dynamics in various brain regions.

Methods Used

  • The primary method involves custom-built glass optrodes and surgery for craniotomies over the target brain regions.
  • The biological model is VGAT channelrhodopsin-II transgenic mice.
  • Electrophysiological recordings and optogenetic stimulation are critical steps in the process.
  • Steps include careful surgical preparation, anesthesia management, and precise insertion techniques for the optrode.
  • Special attention is paid to sterilization and recovery protocols post-surgery.

Main Results

  • The technique allowed well-isolated single-unit activity recording from specific neuron types in the inferior colliculus.
  • Light stimuli resulted in varied responses, exciting GABAergic neurons and suppressing glutamatergic neurons.
  • These findings suggest the method's efficacy in analyzing neural circuitry and neuron-type specific responses.
  • Crucially, it enables detailed investigation into neuron dynamics in awake and behaving mice.

Conclusions

  • This study establishes a reliable optogenetic recording method that enhances neuroscience research capabilities.
  • The technique significantly aids in comprehending the interaction and functional roles of different neurons.
  • Overall, it opens avenues for deeper exploration of neuron mechanisms and interactions within various brain regions.

Frequently Asked Questions

What are the advantages of this optogenetic recording method?
This method allows for reliable single-unit recordings from awake mice, providing insights into neuron functionality with cost-effective materials.
How is the biological model utilized in the experiment?
The experiment utilizes VGAT channelrhodopsin-II transgenic mice to study GABAergic neuron responses to light stimuli.
What types of data or outcomes are obtained using this method?
The method yields detailed electrophysiological data and allows for the assessment of neuron-type specific responses during light stimulation.
How can this optogenetic method be applied in research?
Researchers can employ this technique to explore neuron interactions, plasticity, and functional roles in various neurological contexts.
What should be considered when implementing this technique?
Proper surgical techniques and recovery protocols are essential to ensure the success of the recordings and the welfare of the subjects.
What limitations does this study acknowledge?
One limitation may include the complexity of the surgical procedure, requiring careful execution to minimize stress or injury to the subject.
How does this method contribute to understanding neuronal mechanisms?
It enables investigations into the activities and responses of different neuron types in real time, improving our understanding of neural circuitry.

Ce travail présente une méthode pour exécuter une seule unité optogenetic enregistrement fiable d’une souris éveillée à l’aide d’une optrode de verre sur mesure.

Cette méthode peut aider à répondre à des questions clés dans le domaine des neurosciences, telles que les fonctions de différents types de neurones. Le principal avantage de cette technique est qu’elle permet d’identifier de manière fiable les différents types de neurones in vivo avec des matériaux peu coûteux et accessibles. Je ferai une démonstration de la procédure avec Shinji Muramoto, notre technicien, et Lanlan Ma, notre professeur adjoint de notre laboratoire.

Pour commencer cette procédure, retirez doucement le tube d’acier pour le contrôle de la pression du canon du support. Élargissez le trou du côté du siège de la goupille dans le canon du support en perçant. Ensuite, placez un tuyau en acier inoxydable dans le trou.

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