Stereotaxic Surgery for Genetic Manipulation in Striatal Cells of Neonatal Mouse Brains

Shih-Yun Chen; Hsiao-Ying Kuo; Fu-Chin Liu

doi:10.3791/57270

Stereotaxic Surgery for Genetic Manipulation in Striatal Cells of Neonatal Mouse Brains

JoVE Journal
Neuroscience

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JoVE Journal Neuroscience

Stereotaxic Surgery for Genetic Manipulation in Striatal Cells of Neonatal Mouse Brains

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09:44 min

July 10, 2018

DOI: 10.3791/57270-v

Shih-Yun Chen¹, Hsiao-Ying Kuo¹, Fu-Chin Liu^1,2

¹Institute of Neuroscience,National Yang-Ming University, ²Brain Research Center,National Yang-Ming University

Overview

This article outlines a stereotaxic surgery protocol for microinjecting reagents into the striatum of neonatal mouse brains using a homemade head-fixed device. This technique is particularly beneficial for genetic manipulation of neuronal cells within specific brain regions during neonatal development.

Key Study Components

Area of Science

Neuroscience
Genetic Manipulation
Developmental Biology

Background

Stereotaxic surgery is a critical method for targeting specific brain regions.
Neonatal brain development is vital for understanding genetic influences on neurodevelopment.
Creating a homemade device makes this method more accessible and cost-effective.

Purpose of Study

To develop a straightforward protocol for genetic manipulation in neonatal mice.
To enhance precision in microinjections using a homemade device.
To enable the study of neurodevelopmental processes at pivotal stages.

Methods Used

The primary method involves stereotaxic surgery within a controlled environment.
This study uses neonatal mice as a biological model for observing early genetic modifications.
The protocol includes detailed steps for anesthetizing pups and positioning them accurately for injections.
It emphasizes careful handling of surgical instruments to prevent complications during the procedure.

Main Results

Successful microinjections resulted in widespread expression of GFP and tdTomato in striatal cells, indicating effective genetic manipulation.
Results suggest a successful implementation of Cre loxP mediated recombination within the targeted brain regions.
The study supports the potential for subsequent research on genetic impacts during postnatal brain development.

Conclusions

This study demonstrates a feasible approach for conducting targeted genetic injections during early brain development.
The method is accessible and can be adapted for various experimental needs in neuroscience.
It underscores the importance of precise methodologies in understanding neuronal mechanisms and developmental biology.

Frequently Asked Questions

What are the advantages of using a homemade device for surgeries?

A homemade device can be more cost-effective and customizable, allowing for greater accessibility and flexibility in experimental designs.

How is the neonatal pup prepared for injection?

The pup is anesthetized using hypothermia before being positioned in a head tray secured to the stereotaxic apparatus.

What types of outcomes can be measured after the procedure?

Key outcomes include gene expression patterns and potential alterations in neurodevelopment as observed through imaging and molecular assays.

Can this method be adapted for other types of injections?

Yes, the protocol can be modified for different reagents or therapeutic agents, depending on the experimental goals.

What are some limitations of this technique?

Challenges include ensuring accurate targeting during injections and the potential for variability in response based on handling and surgical precision.

How does this method facilitate studying genetic influences in development?

By allowing for precise genetic modifications in specific brain regions, researchers can investigate the roles of targeted genes in neurodevelopment.

We describe a protocol of stereotaxic surgery with a homemade head-fixed device for microinjecting reagents into the striatum of neonatal mouse brains. This technique allows genetic manipulation in neuronal cells of specific regions of neonatal mouse brains.

This method can help answer key questions in the genetic and neuro development such as during postnatal development. The main advantage of this technique is that it is a simple technique with home-made device. Visual demonstration of this method is critical because making targeted injections into the brain requires full attention to detail.

To begin, make a holder to use neonatal pups in a stereotaxic apparatus. First make the head tray. Cut off about 1/5 of the bottom of a 1.5 milliliter centrifuge tube to make an opening for the head of a neonatal pup.

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