Assessment of Long-term Depression Induction in Adult Cerebellar Slices

Kazuhiko Yamaguchi; Masao Ito

doi:10.3791/59859

Évaluation de l'induction à long terme de dépression dans les tranches de cérévelateur adultes

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Neuroscience

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JoVE Journal Neuroscience

Assessment of Long-term Depression Induction in Adult Cerebellar Slices

Évaluation de l'induction à long terme de dépression dans les tranches de cérévelateur adultes

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09:30 min

October 16, 2019

DOI: 10.3791/59859-v

Kazuhiko Yamaguchi¹, Masao Ito²

¹Laboratory for Behavioral Genetics, Center for Brain Science,RIKEN, ²Senior Adviser's Office, Center for Brain Science,RIKEN

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Overview

This study investigates the induction of long-term depression (LTD) in cerebellar Purkinje cells using multiple protocols, especially in gene-manipulated animals. The research highlights a discrepancy between LTD and motor learning, revealing the need for varied approaches to assess LTD effectively in genetic contexts.

Key Study Components

Area of Science

Neuroscience
Electrophysiology
Motor Learning

Background

Gene manipulation can affect neuronal functions.
Long-term depression (LTD) is crucial for synaptic plasticity.
The role of LTD in motor learning is complex and requires clarification.
Multiple experimental protocols are necessary for proper assessment of LTD.

Purpose of Study

To assess LTD induction in cerebellar Purkinje cells.
To explore the relationship between LTD and motor learning.
To apply different protocols to gene-manipulated animal models.

Methods Used

Used ex vivo brain slices from cerebellar Purkinje cells.
Focused on gene-manipulated animal models to study neuronal functions.
Protocols involved chilling the samples, surgical procedures, and electrophysiological recording techniques.
Careful steps were taken to induce LTD using stimuli and whole-cell patch clamp methods.

Main Results

Induction of LTD was demonstrated through various stimulation protocols.
Electrophysiological characteristics like paired pulse facilitation and depression were observed.
Complex spikes in Purkinje cells showed similar patterns in various stimulation conditions.
The study underscores the necessity of adapting protocols to achieve consistent results in genetic manipulation contexts.

Conclusions

This research provides insights into the mechanisms underlying LTD in Purkinje cells.
It emphasizes the importance of experimental variability in understanding neuronal plasticity.
Findings advance knowledge on LTD's role in motor learning and neuronal signaling.

Frequently Asked Questions

What are the advantages of using gene-manipulated animals?

Gene-manipulated animals allow researchers to study specific neuronal functions and plasticity changes at a molecular level, making it easier to explore causal relationships in motor behavior.

How is LTD induced in cerebellar Purkinje cells?

LTD is induced through a combination of parallel fiber and climbing fiber stimulations, monitored by electrophysiological techniques, such as whole-cell patch clamping.

What types of data are obtained in this study?

Data collected include electrophysiological responses, input resistance changes, and characteristics of synaptic facilitation or depression, providing insights into neuronal signaling.

What are the main limitations of this research?

The study may face limitations related to the specificity of gene manipulations and the variability inherent in physiological responses across different protocols.

How can the methods used in this study be adapted for other models?

Methodological approaches can be adapted by varying stimulus protocols or selecting different neuronal types for investigating LTD and related plasticity mechanisms.

What is the significance of complex spikes observed in Purkinje cells?

The complex spikes indicate polysynaptic influences and alterations in synaptic connectivity, which are key for understanding neuronal integration and motor learning processes.

Chez certains animaux manipulés par des gènes, l'utilisation d'un seul protocole peut ne pas induire la Ltd dans les cellules cérévelaires de Purkinje, et il peut y avoir un écart entre la LTD et l'apprentissage moteur. Plusieurs protocoles sont nécessaires pour évaluer l'induction de LTD chez les animaux manipulés par des gènes. Les protocoles standard sont affichés.

Lorsqu’une fonction protéique particulière est modifiée par manipulation génétique, la coordination de l’échec irritant et de l’affaiblissement de fonctionnement moteur est considérée comme une condition nécessaire à la relation causale entre l’ILD et l’apprentissage moteur. Ici, nous démontrons l’utilisation de protocoles multiples pour évaluer ltd qui peuvent être induits par des mécanismes compensatoires chez les animaux manipulés par les gènes. Avant de récolter le cerveau, réfrigérer et oxygéner deux beakers de 50 millilitres d’ACSF sur la glace.

Lorsque la température de la solution atteint moins de quatre degrés Celsius, ajouter 50 microlitres d’une tétrodotoxine millimolaire à l’un des beakers glacés. Pour récolter le cerveau, tenez la tête et utilisez des ciseaux ophtalmologiques pour couper la peau superficielle le long de la ligne médiane. Rétractez manuellement la peau pour exposer largement la surface du crâne et utilisez les ciseaux pour couper le long du crâne horizontalement du trou spinocerebellar majeur à juste au-dessus des yeux et des oreilles avant de couper le crâne le long d’une ligne au-dessus des deux yeux.

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