Cortex-, Hippocampus-, Thalamus-, Hypothalamus-, Lateral Septal Nucleus- and Striatum-specific <em>In Utero</em> Electroporation in the C57BL/6 Mouse

Jan Baumgart; Nadine Baumgart

doi:10.3791/53303

Específica Striatum Cortex, hipocampo, Thalamus-, hipotálamo, núcleo-septal lateral y In Uter...

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Neuroscience

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JoVE Journal Neuroscience

Cortex-, Hippocampus-, Thalamus-, Hypothalamus-, Lateral Septal Nucleus- and Striatum-specific In Utero Electroporation in the C57BL/6 Mouse

Específica Striatum Cortex, hipocampo, Thalamus-, hipotálamo, núcleo-septal lateral y In Utero Electroporación en el ratón C57BL / 6

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06:00 min

January 18, 2016

DOI: 10.3791/53303-v

Jan Baumgart¹, Nadine Baumgart¹

¹TARC (Translational Animal Research Center),University Medical Center, JGU Mainz

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Overview

This protocol details the in utero electroporation technique for transfecting specific regions of the C57BL/6 central nervous system. It focuses on areas such as the cortex, hippocampus, thalamus, hypothalamus, lateral septal nucleus, and striatum.

Key Study Components

Area of Science

Neuroscience
Cell Biology
Genetic Engineering

Background

In vivo cell manipulation is crucial for understanding brain development and function.
This method allows for efficient cell type manipulation without generating genetically modified mice.
Electroporation requires precise electrode positioning for successful transfection.
Understanding signaling cascades and cell differentiation is essential for neuroscience research.

Purpose of Study

To modify specific cells in the central nervous system of mice.
To investigate brain development and functionality.
To facilitate research on signaling pathways and cell differentiation.

Methods Used

In utero electroporation technique for transfection.
Use of anesthetized pregnant mice for the procedure.
Application of a saline solution to prevent drying of the abdominal cavity.
Identification of critical electrode positioning for effective electroporation.

Main Results

Successful transfection of targeted brain regions.
Improved understanding of cell signaling and differentiation.
Demonstration of the method's efficiency compared to traditional genetic modification.
Guidance for new researchers on electrode positioning.

Conclusions

The in utero electroporation technique is effective for CNS manipulation.
This method enhances research capabilities in neuroscience.
It provides a faster alternative to generating genetically modified mice.

Frequently Asked Questions

What is in utero electroporation?

In utero electroporation is a technique used to introduce DNA into specific regions of the developing central nervous system in embryos.

What are the advantages of this method?

It allows for rapid and efficient manipulation of cells without the need for creating genetically modified mice.

Which brain regions can be targeted?

The cortex, hippocampus, thalamus, hypothalamus, lateral septal nucleus, and striatum can be targeted using this technique.

How does this method help in neuroscience research?

It aids in understanding brain development, cell signaling pathways, and the differentiation of various cell types.

What precautions should be taken during the procedure?

It is important to keep the abdominal cavity moist and ensure precise electrode positioning for successful electroporation.

Is this method suitable for beginners?

Yes, but it requires careful attention to technique, especially in electrode placement.

Este protocolo describe en detalle cómo transfectar específicamente diferentes regiones en el sistema nervioso central C57BL/6 a través de la electroporación en el útero. En este protocolo se incluyen instrucciones detalladas para las transfecciones de regiones que se desarrollan en la corteza, el hipocampo, el tálamo, el hipotálamo, el núcleo septal lateral y el cuerpo estriado.

El objetivo general de esta manipulación celular in vivo es modificar específicamente las células de ciertas regiones del sistema nervioso central de la corteza similar a la del ratón, el hipocampo, el tálamo, el hipotálamo, el núcleo septal lateral y el cuerpo estriado. Este método puede ayudar a responder preguntas clave sobre el desarrollo y la función del cerebro, por ejemplo, a través de la identificación de diferentes cascadas de señalización y la determinación del nivel de diferenciación celular. La principal ventaja de esta técnica es que permite la manipulación rápida y eficiente de varios tipos de células en el sistema nervioso central sin la generación de ratones modificados genéticamente, que requiere mucho tiempo.

En general, la demostración de este método es útil, porque las personas nuevas en este método a menudo tendrán dificultades para encontrar la posición correcta del electrodo, que es fundamental para una electroporación específica. Para comenzar este procedimiento, abra la cavidad abdominal de una rata embarazada anestesiada con un bisturí. Evite que la cavidad abdominal abierta se seque humedeciéndola con una solución salina de alcohol metílico tibia al 0,9%.

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Neurociencia Número 107 En el útero Electroporación ángulo-mapa C57BL / 6 el sistema nervioso central la administración dirigida de genes corteza el hipocampo el tálamo el hipotálamo núcleo septal lateral cuerpo estriado