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Neuroscience
嗅球的四次免疫染色用于嗅觉感觉轴突分子识别码的可视化
嗅球的四次免疫染色用于嗅觉感觉轴突分子识别码的可视化
JoVE Journal
Neuroscience
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JoVE Journal Neuroscience
Quadruple Immunostaining of the Olfactory Bulb for Visualization of Olfactory Sensory Axon Molecular Identity Codes

嗅球的四次免疫染色用于嗅觉感觉轴突分子识别码的可视化

Full Text
7,625 Views
06:32 min
June 5, 2017

DOI: 10.3791/55893-v

Naoki Ihara1, Yuji Ikegaya1,2, Haruki Takeuchi1,3

1Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences,The University of Tokyo, 2Center for Information and Neural Networks,National Institute of Information and Communications Technology, 3PRESTO,Japan Science and Technology Agency (JST)

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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 protocol outlines an immunohistochemical staining method to visualize the expression of axon-sorting molecules at the axon termini of olfactory sensory neurons. This technique is crucial for understanding olfactory development processes such as axon guidance and synapse formation.

Key Study Components

Area of Science

  • Neuroscience
  • Olfactory development
  • Immunohistochemistry

Background

  • Olfactory sensory neurons play a key role in the sense of smell.
  • Axon-sorting molecules are essential for establishing proper neural circuitry.
  • Understanding their expression patterns can provide insights into olfactory system functions.
  • Visualizing these expressions can aid in studying regeneration processes in olfactory sensory neurons.

Purpose of Study

  • To visualize the combined expression of axon-sorting molecules.
  • To characterize key processes in olfactory development.
  • To facilitate comparison of expression patterns in the olfactory bulb.

Methods Used

  • Immunohistochemical staining technique.
  • Use of mouse heads aged one to two weeks.
  • Intracardiac perfusion with 4% paraformaldehyde for fixation.
  • Embedding and sectioning of olfactory tissue samples.

Main Results

  • Successful visualization of axon-sorting molecule expressions.
  • Demonstrated the ability to analyze expression patterns without section variation.
  • Provided a clear method for embedding olfactory tissues.
  • Highlighted the importance of visual demonstrations for learning the technique.

Conclusions

  • The immunohistochemical method is effective for studying olfactory sensory neurons.
  • Understanding axon-sorting molecules can enhance knowledge of olfactory system development.
  • This technique can be a valuable tool for future research in olfactory regeneration.

Frequently Asked Questions

What is the main goal of this study?
The main goal is to visualize the expression of axon-sorting molecules in olfactory sensory neurons.
Why is this method important?
It allows for the analysis of expression patterns without variation between sections, aiding in the understanding of olfactory development.
What age of mice is used in this protocol?
Mice aged one to two weeks are used for the study.
What fixation method is employed?
Intracardiac perfusion with 4% paraformaldehyde is used for fixation.
What challenges are associated with this technique?
The embedding step can be difficult to learn due to the positioning of the olfactory tissue samples.
How does this method contribute to olfactory research?
It provides insights into axon guidance, synapse formation, and regeneration in the olfactory system.

嗅觉神经元表达各种各样的轴突分选分子,建立适当的神经电路。该方案描述了一种免疫组织化学染色方法来观察嗅觉神经元轴突末端的轴突分选分子的组合表达。

这种免疫组织化学染色方法的总体目标是可视化嗅觉感觉神经元轴突末端轴突分选分子的组合表达。这种方法可以帮助表征嗅觉发育领域的关键过程,例如轴突导向、突触形成和嗅觉感觉神经元的再生。这项技术的主要优点是它允许科学家比较和分析嗅球中轴突分选分子的表达模式,而无需维持切片之间的差异。

这种方法的视觉演示至关重要,因为包埋步骤很难学习,因为嗅觉组织样本在模具中的位置和角度很难用语言描述。从 1 到 2 周龄的小鼠的头部开始,这些小鼠通过心内灌注 4% 多聚甲醛固定。将剪刀刀片插入上下牙齿之间的空间,水平切割以去除下颌骨。

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