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JoVE Journal
Neuroscience
Imagem de lapso de tempo de neurônios migratórios e progenitores gliais em fatias de cérebro de c...
Imagem de lapso de tempo de neurônios migratórios e progenitores gliais em fatias de cérebro de c...
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Time-Lapse Imaging of Migrating Neurons and Glial Progenitors in Embryonic Mouse Brain Slices

Imagem de lapso de tempo de neurônios migratórios e progenitores gliais em fatias de cérebro de camundongo embrionário

Full Text
1,590 Views
04:17 min
March 8, 2024

DOI: 10.3791/66631-v

Hidenori Tabata1,2, 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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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.

Durante o desenvolvimento do córtex cerebral, os neurônios e as células gliais se originam na zona ventricular que reveste o ventrículo e migram em direção à superfície do cérebro. Muitos genes estão envolvidos neste processo. Este protocolo apresenta a técnica para a imagem de lapso de tempo de neurônios migratórios e progenitores gliais.

Durante o desenvolvimento do córtex cerebral, neurônios e células cinzentas são derivados da zona ventricular e migram para outra parte da superfície do cérebro. Muitos genes estão envolvidos nesse processo, incluindo aqueles responsáveis por distúrbios do neurodesenvolvimento e psiquiátricos. Estamos abordando suas funções nos três comportamentos neste processo.

Recentemente, relatamos que os progenitores de astrócitos assumem dois modos distintos de migração: migração errática e guiada por vasos sanguíneos. Essas observações foram feitas usando uma combinação de rotulagem específica do sorotipo e métodos de observação de tempo apresentados neste vídeo. Para nivelar as células, utilizamos o sistema de eletroporação no útero, que desenvolvemos para visualizar células individuais com uma alta relação sinal-ruído.

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Keywords: Imagem de lapso de tempo migração de neurônios migração de progenitores gliais fatias de cérebro de camundongo embrionário desenvolvimento do córtex cerebral zona ventricular superfície cerebral distúrbios do neurodesenvolvimento distúrbios psiquiátricos eletroporação in utero marcação fluorescente de proteínas manipulação gênica comportamento migratório velocidade de migração imagem de células vivas

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