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JoVE Journal
Neuroscience
Whole-mount Imaging of Mouse Embryo Sensory Axon Projections
Whole-mount Imaging of Mouse Embryo Sensory Axon Projections
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Whole-mount Imaging of Mouse Embryo Sensory Axon Projections

Whole-mount Imaging of Mouse Embryo Sensory Axon Projections

Full Text
10,508 Views
08:37 min
December 9, 2014

DOI: 10.3791/52212-v

Kevin J. O’Donovan1, Catherine O’Keeffe1, Jian Zhong1

1Brain and Mind Research Institute, Burke Medical Research Institute,Weill Medical College of Cornell University

Overview

This article presents an optimized protocol for genotyping, staining, and preparing fetal mice to visualize peripheral nociceptor axon projections. This method effectively assesses sensory axon growth phenotypes in developing genetically engineered mice.

Key Study Components

Area of Science

  • Neuroscience
  • Developmental Biology
  • Genetics

Background

  • Understanding sensory axon growth is crucial for insights into pain mechanisms.
  • Genetically engineered mice provide a model for studying these processes.
  • Visualization of axon projections is essential for assessing growth phenotypes.
  • Existing methods may not adequately reveal deep-lying axon projections.

Purpose of Study

  • To develop a reliable protocol for assessing sensory axon growth in fetal mice.
  • To enable visualization of nociceptive axon projections in whole embryos.
  • To facilitate the screening of genetic modifications on sensory axon growth.

Methods Used

  • Breeding and genotyping of tre a tau laxy reporter mice.
  • Fixation and staining of mouse embryos for visualization.
  • Dehydration and clearing of embryos to enhance visibility of axon projections.
  • Utilization of the track a Tau Lae reporter for assessing growth phenotypes.

Main Results

  • The protocol effectively visualizes sensory axon projections in fetal mice.
  • Results demonstrate rapid assessment of embryonic nociceptive axon growth.
  • Allows researchers to investigate changes in sensory axon growth due to genetic modifications.
  • Provides a framework for future studies in sensory axon development.

Conclusions

  • This optimized protocol is a valuable tool for studying sensory axon growth.
  • It enhances the ability to visualize complex axon projections in embryos.
  • The method supports further research into genetic influences on sensory development.

Frequently Asked Questions

What is the main goal of this study?
The main goal is to assess sensory axon growth phenotypes in developing fetal mice.
How are the embryos prepared for imaging?
Embryos are fixed, stained, dehydrated, and cleared for visualization.
What type of mice are used in this study?
Tre a tau laxy reporter mice are used for genotyping and imaging.
What does the track a Tau Lae reporter do?
It helps visualize sensory axon growth and assess genetic modifications.
Why is this method important?
It allows for rapid assessment of nociceptive axon growth in embryos, aiding research.
Can this method be applied to other genetic backgrounds?
Yes, researchers can cross their gene of interest onto the track a tlac Z background.

We present here an optimized protocol to genotype, stain and prepare fetal mice for the imaging of peripheral nociceptor axon projections in the whole animal, as an effective method to assess sensory axon growth phenotypes in developing genetically engineered mice.

The overall goal of the following experiment is to effectively assess sensory axon growth phenotypes in developing fetal mice. This is achieved by first properly breeding and genotyping tre a tau laxy reporter mice such that downstream staining assays can be performed as a second step. The mouse embryos are fixed and stained to allow for visualization of sensory axon projections in the whole animal.

Next, the embryos are dehydrated and cleared in order to completely visualize the deep lying axon projections that would otherwise not be visible. The results demonstrate that these methods are an effective means of rapidly assessing embryonic nociceptive axon growth phenotypes. Using the track a Tau Lae reporter, This method can help answer key questions in the field of sensory axon growth by allowing researchers to cross their gene of interest onto the track a tlac Z background and screen for any changes to sensory axon growth phenotypes.

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