Measurement of Oxygen Consumption Rate in Acute Striatal Slices from Adult Mice

Lianteng Zhi; Jingyu Zhao; David Jaffe; Yuanxin Chen; Ninghan Wang; Qi Qin; Erin L. Seifert; Chenjian Li; Hui Zhang

doi:10.3791/63379

Mesure du taux de consommation d’oxygène dans les tranches striatales aiguës de souris adultes

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Neuroscience

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JoVE Journal Neuroscience

Measurement of Oxygen Consumption Rate in Acute Striatal Slices from Adult Mice

Mesure du taux de consommation d’oxygène dans les tranches striatales aiguës de souris adultes

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07:41 min

June 8, 2022

DOI: 10.3791/63379-v

Lianteng Zhi¹, Jingyu Zhao¹, David Jaffe¹, Yuanxin Chen¹, Ninghan Wang^1,2, Qi Qin¹, Erin L. Seifert³, Chenjian Li⁴, Hui Zhang¹

¹Department of Neuroscience,Thomas Jefferson University, ²School of Biomedical Engineering,Drexel University, ³Department of Pathology, MitoCare Center,Thomas Jefferson University, ⁴School of Life Sciences,Peking University

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Overview

This study presents a novel method for measuring oxygen consumption rate (OCR) in acute striatal slices from adult mice using a Seahorse XF analyzer. This approach aims to provide a more physiologically relevant assessment of mitochondrial function compared to traditional methods, especially for disease models like Parkinson's and Huntington's diseases.

Key Study Components

Area of Science

Neuroscience
Mitochondrial function
Disease modeling

Background

Oxygen consumption rate is an established indicator of mitochondrial function.
Acute striatal slices closely approximate physiological conditions.
Current methods may lack relevance to in vivo cellular environments.
The technique may enhance understanding of neurodegenerative diseases.

Purpose of Study

To develop a more physiologically relevant method for measuring OCR.
To investigate mitochondrial function in acute brain slices.
To support research into disease mechanisms in neurodegenerative disorders.

Methods Used

Utilized Seahorse XF analyzer for OCR measurement.
Focused on acute striatal slices from adult mice.
Provided protocols for slice preparation, incubation, and measurement.
Highlighted calibration and setup steps for the assay.

Main Results

Stable basal respiration rates were observed across different slice conditions.
OCR measurements demonstrated proportional relationships to slice volume.
Mitochondrial coupling efficiency was highest at specific slice thickness and punch size.
Differential OCR findings were noted between knockout and wild-type mice.

Conclusions

This method enhances the ability to study mitochondrial function in a biologically relevant context.
May lead to better insights into neurodegenerative disease mechanisms.
Supports future research and therapeutic strategies in mitochondrial-related disorders.

Frequently Asked Questions

What are the advantages of using acute striatal slices?

Acute striatal slices provide a more accurate representation of the physiological environment compared to isolated mitochondria or cultured cells, allowing for better assessment of cellular function.

What types of data can be obtained through this method?

The method allows for direct measurements of oxygen consumption rates, which are indicative of mitochondrial function and can reveal insights into cellular metabolism in the context of disease.

How is the sample preparation process conducted?

The preparation involves delicately dissecting brain tissue, slicing it to a specified thickness, and incubating it in a respiration buffer prior to OCR measurement.

Can this method be adapted to study other brain regions?

The methodology can potentially be applied to other brain regions, allowing researchers to explore mitochondrial function across various neuronal environments.

What limitations should researchers be aware of?

Factors such as slice thickness, incubation conditions, and the age of the mice can influence the results and should be carefully controlled for consistency.

How does this research impact our understanding of neurodegenerative diseases?

By enabling more physiologically relevant assessments, this research can help elucidate mitochondrial dysfunctions associated with neurodegenerative diseases like Parkinson's and Huntington's disease.

Le taux de consommation d’oxygène (OCR) est un indicateur commun de la fonction mitochondriale et peut être utilisé pour étudier différents modèles de maladie. Nous avons développé une nouvelle méthode utilisant un analyseur Seahorse XF pour mesurer directement l’OCR dans les tranches striatales aiguës de souris adultes qui est plus pertinente physiologiquement que d’autres méthodes.

Nous avons développé une nouvelle méthode utilisant un analyseur XF hippocampe pour mesurer directement le taux de consommation d’oxygène. Mais proxy commun pour la fonction mitochondriale dans la tranche de lame aiguë de souris adultes. Cette méthode mesure un cellulaire par l’énergie dans les ponctions à partir de structures cérébrales anatomiquement définies.

L’utilisation de tranches aiguës imite plus étroitement l’environnement cellulaire physiologique, ce qui ne peut pas être réalisé de la même manière que les cellules mitochondriales ou de culture isolées. Cette méthode intéressera grandement les chercheurs travaillant dans le domaine de la maladie de Parkinson et de la maladie de Huntington. Pour commencer, ouvrez le kit de dosage de flux extracellulaire et retirez à la fois la cartouche du capteur et la plaque utilitaire.

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