Time-Lapse Imaging of Migrating Neurons and Glial Progenitors in Embryonic Mouse Brain Slices

Hidenori Tabata; Koh-ichi Nagata; Kazunori Nakajima

doi:10.3791/66631

マウス胚脳切片における移動するニューロンとグリア前駆細胞のタイムラプスイメージング

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Neuroscience

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JoVE Journal Neuroscience

Time-Lapse Imaging of Migrating Neurons and Glial Progenitors in Embryonic Mouse Brain Slices

マウス胚脳切片における移動するニューロンとグリア前駆細胞のタイムラプスイメージング

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04:17 min

March 8, 2024

DOI: 10.3791/66631-v

Hidenori Tabata^1,2, Koh-ichi Nagata², Kazunori Nakajima¹

¹Department of Anatomy,Keio University School of Medicine, ²Department of Molecular Neurobiology, Institute for Developmental Research,Aichi Developmental Disability Center

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Overview

This study focuses on the development of the cerebral cortex, highlighting the migration of neurons and glial progenitors from the ventricular zone to the brain surface. By employing time-lapse imaging techniques alongside in utero electroporation for cell labeling, the research investigates the migration modes of astrocyte progenitors and the genes influencing neurodevelopmental and psychiatric disorders.

Key Study Components

Area of Science

Neurodevelopment
Cell migration
Neurobiology

Background

The study explores the origin of neurons and glial cells during cortical development.
It addresses the role of specific genes in neurodevelopmental and psychiatric disorders.
Astrocyte progenitors exhibit distinct migration modes relevant to cortical formation.
In vivo gene transfer techniques facilitate the observation of these migration patterns.

Purpose of Study

To visualize and analyze the behaviors of migrating neurons and glial cells.
To elucidate the functional roles of genes involved in neurodevelopment.
To investigate cell behavior and interactions between neurons, glia, and blood vessels.

Methods Used

Utilized in utero electroporation for gene transfer and cell labeling.
Focused on mouse models to examine neuronal and glial progenitor migration.
Involved time-lapse imaging of EGFP- and RFP-positive cells.
The protocol outlined specific surgical and imaging techniques for accurate observations.
Included the creation of brain sections for analysis via a vibrating microtome.

Main Results

Identified two distinct astrocyte progenitor migration modes: erratic and blood vessel-guided.
Presented data showed defined trajectories of migrating cells over time.
Findings contribute to the understanding of neurodevelopmental disorder mechanisms.
Demonstrated the effectiveness of electroporation for visualizing individual cells.

Conclusions

This study provides insights into the mechanisms of neuronal migration during cortical development.
Findings may enhance our understanding of neurodevelopmental and psychiatric disorders.
The results underscore the complex interplay between glial and vascular components in brain development.

Frequently Asked Questions

What are the advantages of using in utero electroporation?

In utero electroporation allows for precise gene transfer and labeling of specific cells in vivo, providing high signal-to-noise ratios essential for imaging individual cells.

How does the experimental model contribute to the understanding of CNS development?

The use of mouse models enables the observation of real-time neuronal migration and the effects of genetic variations, offering insights into brain development processes.

What types of data can be obtained from this time-lapse imaging method?

The method provides quantitative data on migrating cell trajectories, enabling the analysis of different migration modes and their dynamics over time.

Can this method be adapted for other cell types or conditions?

Yes, the protocol can be modified to study various cell types or conditions, making it a versatile tool for examining different aspects of cell behavior in vivo.

What are some limitations of the approach used in this study?

Potential limitations include the complexity of in utero manipulation and the specificity of gene delivery, which may affect reproducibility across different experiments.

What cellular behaviors were focused on in this research?

The study concentrated on the migration patterns of neurons and glial cells, specifically investigating how astrocyte progenitors navigate during cortical development.

大脳皮質の発達中、ニューロンとグリア細胞は、心室の内側を覆う心室ゾーンで発生し、脳表面に向かって移動します。このプロセスには多くの遺伝子が関与しています。このプロトコルでは、移動するニューロンとグリア前駆細胞のタイムラプスイメージングの技術を紹介します。

大脳皮質の発達中、ニューロンと灰色細胞は心室領域に由来し、脳表面の他の部分に移動します。このプロセスには、神経発達障害や精神障害の原因となる遺伝子など、多くの遺伝子が関与しています。このプロセスにおける3つの振る舞いについて、それらの機能に取り組んでいます。

最近、アストロサイト前駆細胞は、不規則な移動と血管誘導移動という2つの異なる移動モードをとることが報告されています。これらの観察は、このビデオで紹介した血清型特異的な標識法と時間観察法の組み合わせを使用して行われました。細胞の平準化には、S/N比の高い個々の細胞を可視化するために開発した子宮内エレクトロポレーションシステムを利用しています。

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