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Neuroscience
Corte cerebral de ratón agudo para investigar la actividad espontánea de la red de hipocampo
Corte cerebral de ratón agudo para investigar la actividad espontánea de la red de hipocampo
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Acute Mouse Brain Slicing to Investigate Spontaneous Hippocampal Network Activity

Corte cerebral de ratón agudo para investigar la actividad espontánea de la red de hipocampo

Full Text
10,412 Views
07:58 min
August 28, 2020

DOI: 10.3791/61704-v

Alexander C. Whitebirch1

1Department of Neuroscience, Mortimer B. Zuckerman Mind Brain Behavior Institute,Columbia 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 details a protocol for preparing horizontal hippocampal-entorhinal cortex slices from mice, enabling investigation of network oscillations in acute brain slice preparations. The method emphasizes maintaining spontaneous activity through specific incubation and recording conditions using artificial cerebrospinal fluid.

Key Study Components

Area of Science

  • Neuroscience
  • Electrophysiology
  • Neurobiology

Background

  • Horizontal hippocampal-entorhinal cortex slices are critical for studying neural network activity.
  • Study of sharp-wave ripple activity provides insights into hippocampal function.
  • Pharmacological and optogenetic manipulations can be performed in this model.
  • Spontaneous network oscillations are vital for understanding neural communication.

Purpose of Study

  • To investigate mechanisms underlying hippocampal network oscillations.
  • To refine a protocol for generating viable slices that maintain network activity.
  • To enable localized electrophysiological recordings in a controlled environment.

Methods Used

  • The protocol includes the preparation of ex vivo brain slices from mice.
  • It focuses on horizontal hippocampal-entorhinal cortex tissue.
  • No multiomics workflows are mentioned in the study.
  • Slices are incubated under precise conditions to promote spontaneous activity.
  • The timing for recovery and the setup required for electrophysiological recordings are detailed.

Main Results

  • The slices exhibit spontaneous sharp-wave ripple activity under the specified conditions.
  • Electrical stimulation leads to characteristic field post-synaptic potentials, indicating healthy slices.
  • In suboptimal conditions, altered electrophysiological responses are observed.
  • The findings validate the protocol's effectiveness for studying local field potentials in a dynamic neuronal network.

Conclusions

  • This study demonstrates a robust protocol for preparing and recording from mouse brain slices.
  • The insights gained contribute significantly to understanding hippocampal oscillatory dynamics.
  • The implications extend towards improved investigation of neuronal mechanisms and potential disease models.

Frequently Asked Questions

What are the advantages of using horizontal hippocampal slices?
Horizontal hippocampal slices allow for localized electrophysiological recordings and maintenance of spontaneous neural activity, providing insights into network dynamics.
How is the biological model implemented in this study?
The model involves preparing slices from the hippocampal-entorhinal cortex of mice, enabling researchers to study network-level oscillations under controlled conditions.
What types of data are obtained from this procedure?
Electrophysiological data such as local field potentials and responses to electrical stimulation are collected, reflecting neuronal health and activity.
Can this method be adapted for different neural circuits?
Yes, the protocol can be modified for various brain regions or conditions by adjusting dissection and incubation parameters.
What are some limitations of this approach?
Potential limitations include variability in slice quality and the need for precise environmental control during experiments.
How can this research inform potential therapeutic approaches?
Understanding oscillatory dynamics in the hippocampus may contribute to developing strategies for treating neurological disorders involving network dysfunction.

Este protocolo describe la preparación de rodajas horizontales de corteza hipocampal-entorrinaal (HEC) de ratones que exhiben actividad espontánea de ondulación de onda aguda. Las rodajas se incuban en una cámara de retención de interfaz simplificada y las grabaciones se realizan en condiciones sumergidas con líquido cefalorraquídeo artificial de flujo rápido para promover la oxigenación tisular y la aparición espontánea de la actividad a nivel de red.

Con este protocolo, los investigadores pueden investigar los mecanismos subyacentes a las oscilaciones de la red del hipocampo en preparaciones agudas de cortes de cerebro de ratón. En esta preparación, los registros se realizan en condiciones sumergidas en cortes generando oscilaciones espontáneas de la red, lo que permite realizar técnicas farmacológicas y optogenéticas, y visualizar registros de una sola célula. Para la perfusión transcárdica, inmediatamente antes de retirar el ratón de la cámara de isoflurano, llene la tapa de una placa de Petri con tres a cinco milímetros de solución de sacarosa enfriada y llene el fondo de la placa de Petri con aproximadamente un centímetro de solución de sacarosa enfriada.

Transfiera rápidamente el ratón anestesiado a la almohadilla absorbente izquierda y use tres piezas de cinta adhesiva para asegurar las extremidades delanteras y la cola. Con unas pinzas de tejido grandes y unas tijeras quirúrgicas, cubra la piel y haga una incisión longitudinal desde la parte inferior del esternón hasta la parte superior del pecho. Use las pinzas para tirar hacia arriba del esternón y use las tijeras para cortar el diafragma.

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