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

Registros agudos de electrodos múltiples de una sola unidad del tronco encefálico de ratones con ...

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JoVE Journal Neuroscience

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

Registros agudos de electrodos múltiples de una sola unidad del tronco encefálico de ratones con cabeza fija

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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.

Este protocolo describe un método para obtener grabaciones in vivo de una sola neurona de alta densidad a partir del tronco encefálico de ratones con cabeza fija. Este enfoque se implementa para medir el potencial de acción de las neuronas en el gris periacueductal ventrolateral, una región del tronco encefálico inactiva durante el sueño de movimiento ocular rápido (REM), antes y durante la anestesia general.

Nuestro laboratorio investiga los efectos de los anestésicos de uso común en las neuronas relacionadas con el sueño y sus vías de comunicación. Nuestro objetivo es comprender mejor cómo los anestésicos afectan las vías endógenas del sueño y afectan el sueño perioperatorio y la cognición. Recientemente identificamos los sustratos anatómicos de algunos de los cambios del sueño causados por ciertos tipos de anestésicos generales.

Lo hicimos utilizando ratones modificados genéticamente en los que hay expresión endógena de c-Fos, un marcador de activación neuronal. Sin embargo, con esta nueva técnica, podremos medir directamente la excitabilidad celular en grupos de neuronas del tronco encefálico que están asociadas con el sueño. Esta es una gran ventaja sobre la simple cuantificación de la expresión de c-Fos como marcador sustituto de la activación neuronal.

Nuestro próximo paso será investigar los mecanismos celulares y moleculares que subyacen a los cambios en el sueño inducidos por ciertos tipos de anestésicos generales. Esto implicará medir la excitabilidad celular en las neuronas del tronco encefálico asociadas con el sueño utilizando sondas de neuropíxeles, así como el uso de enfoques transcriptómicos espaciales para cuantificar y mapear la expresión de genes relacionados con el sueño en estas áreas.

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