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

Zeitraffer-Bildgebung von migrierenden Neuronen und glialen Vorläuferzellen in embryonalen Hirnsc...

JoVE Journal
Neuroscience

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Neuroscience

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

Zeitraffer-Bildgebung von migrierenden Neuronen und glialen Vorläuferzellen in embryonalen Hirnschnitten von Mäusen

Full Text

1,520 Views

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

Please note that some of the translations on this page are AI generated. Click here for the English version.

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.

Während der Entwicklung der Großhirnrinde stammen Neuronen und Gliazellen aus der ventrikulären Zone, die den Ventrikel auskleidet, und wandern zur Gehirnoberfläche. Viele Gene sind an diesem Prozess beteiligt. Dieses Protokoll stellt die Technik für die Zeitraffer-Bildgebung von migrierenden Neuronen und glialen Vorläuferzellen vor.

Während der Entwicklung der Großhirnrinde stammen Neuronen und graue Zellen aus der ventrikulären Zone und wandern in andere Teile der Gehirnoberfläche. Viele Gene sind an diesem Prozess beteiligt, darunter auch diejenigen, die für neurologische Entwicklungsstörungen und psychiatrische Störungen verantwortlich sind. Wir befassen uns mit ihren Funktionen in Bezug auf die drei Verhaltensweisen in diesem Prozess.

Kürzlich haben wir berichtet, dass Astrozyten-Vorläuferzellen zwei unterschiedliche Migrationsmodi annehmen: Unregelmäßige und blutgefäßgesteuerte Migration. Diese Beobachtungen wurden mit einer Kombination aus serotypspezifischer Markierung und Zeitbeobachtungsmethoden gemacht, die in diesem Video vorgestellt wurden. Um Zellen zu nivellieren, verwenden wir ein In-utero-Elektroporationssystem, das wir entwickelt haben, um einzelne Zellen mit einem hohen Signal-Rausch-Verhältnis sichtbar zu machen.

Dieses in vivo Gentransfersystem ermöglicht es uns auch, auf einfache Weise Funktionsgewinn- oder Funktionsverlustexperimente an den gegebenen Genen durch Elektroporation ihrer Expressions- oder Neutronenvektoren durchzuführen. Mit diesem experimentellen System wollen wir das zelluläre Verhalten von Neuronen, Gliazellen und Blutgefäßen beobachten und die Wechselwirkung zwischen ihnen aufklären. Die Erkenntnisse aus diesen Studien werden dazu beitragen, die Pathogenese von neurologischen Entwicklungsstörungen zu verstehen.

View the full transcript and gain access to thousands of scientific videos