Acute Single-Unit Multi-Electrode Recordings from the Brainstem of Head-Fixed Mice

Magdalena Pikus; Elzbieta Dulko; Mark Beenhakker; Nadia Lunardi

doi:10.3791/67205

JoVE Journal
Neuroscience

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JoVE Journal Neuroscience

Acute Single-Unit Multi-Electrode Recordings from the Brainstem of Head-Fixed Mice

頭部固定マウスの脳幹からの急性単一ユニット多電極記録

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06:37 min

October 11, 2024

DOI: 10.3791/67205-v

Magdalena Pikus¹, Elzbieta Dulko², Mark Beenhakker³, Nadia Lunardi¹

¹Department of Anesthesiology,University of Virginia, ²Neuroscience Graduate Program,University of Virginia, ³Department of Pharmacology,University of Virginia

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Overview

This study presents a protocol for obtaining in vivo high-density single-neuron recordings from the brainstem of head-fixed mice. The methodology is utilized to assess the action potential firing of neurons in the ventrolateral periaqueductal gray, particularly during general anesthesia and its effects on sleep-related neuron activity.

Key Study Components

Area of Science

Neuroscience
Electrophysiology
Sleep Research

Background

Investigates the impact of anesthetics on neuronal activity.
Focuses on sleep-related neurons and their communication pathways.
Utilizes genetically-modified mice expressing C-fos as a marker of activation.
Seeks to directly measure cellular excitability in sleep-associated neurons.

Purpose of Study

To understand how anesthetics affect endogenous sleep pathways.
To explore cellular and molecular mechanisms of anesthesia-induced sleep changes.
To improve methodologies for measuring neuronal excitability during sleep and anesthesia.

Methods Used

Utilizes high-density silicon probes for electrophysiological recordings.
Employs head-fixed mouse models for precise brainstem targeting.
No multiomics analysis mentioned.
Involves detailed surgical procedures including craniotomy and probe insertion.
Records neuronal firing before and during administration of Sevoflurane anesthesia.

Main Results

The study found a significant decrease in vlPAG neuronal firing during Sevoflurane anesthesia.
Direct measurement of cellular excitability provides insights beyond C-fos expression alone.
Findings support the hypothesis that anesthetics modulate sleep-related neuronal activity.
Results show consistent decreased excitability across all vlPAG neurons assessed.

Conclusions

This method allows for direct investigation of neuronal excitability during anesthesia.
Improved understanding of mechanisms regulating sleep and anesthesia effects.
Implications for future studies on neuronal communications in anesthetic contexts.

Frequently Asked Questions

What advantages does the head-fixed mouse model provide?

The head-fixed model allows for precise targeting of brain regions, facilitating high-density recordings during anesthesia without movement artifacts.

How is the brainstem area identified for recording?

Surgical procedures involve using a mouse-stereotaxic atlas to identify and mark coordinates based on anatomical landmarks like bregma and lambda.

What types of data are obtained from this method?

Data includes action potential firing rates from recorded neurons, assessing changes in excitability before and during anesthesia.

Can this method be adapted for other brain regions?

Yes, the protocol can be adapted for different brain regions by altering the stereotaxic coordinates and surgical approach.

What are the limitations of this protocol?

Limitations include the complexity of the surgical procedures and the potential variability in anesthetic effects on different neuronal populations.

このプロトコルは、頭部固定マウスの脳幹から in vivoで高密度の単一ニューロン記録を取得する方法を説明する。このアプローチは、全身麻酔前および全身麻酔中に、腹外側水道周囲灰色 (急速眼球運動 (REM) 睡眠中に不活性な脳幹領域) のニューロンの活動電位発火を測定するために展開されます。

当研究室では、一般的に使用される麻酔薬が睡眠関連ニューロンとそのコミュニケーション経路に及ぼす影響について研究しています。私たちは、麻酔薬が内因性睡眠経路にどのように影響し、周術期の睡眠と認知に影響を与えるかをよりよく理解することを目指しています。最近、特定の種類の全身麻酔薬によって引き起こされる睡眠変化の一部について、解剖学的基質を特定しました。

これは、ニューロン活性化のマーカーであるc-Fosの内因性発現がある遺伝子改変マウスを使用して行いました。しかし、この新しい技術を使用すると、睡眠に関連する脳幹のニューロンのグループにおける細胞の興奮性を直接測定できるようになります。これは、ニューロン活性化の代理マーカーとしてc-Fos発現を単純に定量化するよりも大きな利点です。

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