Analyzing Dendritic Morphology in Columns and Layers

Chun-Yuan Ting; Philip G. McQueen; Nishith Pandya; Evan S. McCreedy; Matthew McAuliffe; Chi-Hon Lee

doi:10.3791/55410

Analyzing Dendritic Morphology in Columns and Layers

JoVE Journal
Neuroscience

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JoVE Journal Neuroscience

Analyzing Dendritic Morphology in Columns and Layers

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08:41 min

March 23, 2017

DOI: 10.3791/55410-v

Chun-Yuan Ting¹, Philip G. McQueen², Nishith Pandya³, Evan S. McCreedy³, Matthew McAuliffe³, Chi-Hon Lee¹

¹Section on Neuronal Connectivity, Eunice Kennedy Shriver National Institute of Child Health and Human Development,National Institutes of Health (NIH), ²Mathematical and Statistical Computing Laboratory, Center for Information Technology,National Institutes of Health (NIH), ³Biomedical Imaging Research Services Section, Center for Information Technology,National Institutes of Health (NIH)

Overview

This protocol outlines a method to analyze dendritic routing of adult Drosophila medulla neurons in columns and layers. It utilizes dual-view imaging techniques and computational tools to enhance image quality and analyze dendritic structures in 3D space.

Key Study Components

Area of Science

Neuroscience
Neuroanatomy
Imaging Techniques

Background

Dendritic routing is crucial for understanding brain circuitry.
Characterizing dendritic patterns aids in determining cell types.
Identifying mutants can provide insights into neural function.
3D analysis improves comprehension of dendritic wiring logic.

Purpose of Study

To analyze dendritic routing in Drosophila medulla neurons.
To characterize dendritic patterns and determine cell types.
To identify mutants affecting dendritic morphology.

Methods Used

Acquire two image stacks of neurons in orthogonal orientations.
Stain dissected fly brains with specific antibodies.
Label primary antibodies with fluorescent secondary antibodies.
Clear the brain using glycerol and PBS for imaging.

Main Results

Improved imaging techniques enhance dendritic structure analysis.
3D mapping reveals intricate wiring of dendrites.
Characterization of cell types based on morphology is achieved.
Mutant identification provides insights into neural development.

Conclusions

This method significantly advances the understanding of dendritic routing.
It facilitates the mapping of brain circuitry in Drosophila.
Future studies can leverage this technique for broader applications.

Frequently Asked Questions

What is the main goal of this protocol?

The main goal is to analyze dendritic routing of Drosophila medulla neurons.

How does this method improve image quality?

It utilizes a dual-view imaging technique to enhance clarity.

What are the key components used in the staining process?

Rabbit anti-GFP and antibodies that label photoreceptor axons are used.

Why is 3D analysis important in this study?

3D analysis provides a better understanding of dendritic wiring logic.

What insights can be gained from identifying mutants?

Identifying mutants can reveal important aspects of neural function and development.

Here, we show how to analyze dendritic routing of Drosophila medulla neurons in columns and layers. The workflow includes a dual-view imaging technique to improve the image quality and computational tools for tracing, registering dendritic arbors to the reference column array and for analyzing the dendritic structures in 3D space.

The overall goal of this protocol is to analyze dendric routing of adult drosophila medulla neurons in columns and layers to characterize dendric patterns, determine cell types based on morphology, and identify mutants. This method can help answer key questions in the field of neuroscience, including mapping the brain circuitry, and understanding the wiring mechanism of the dendrite. The main advantage of this technique is that it passes down the dendritic properties in three dimensional space, thus improving the understanding of dendritic wiring logic and functions.

The goal of this procedure is to acquire two image stacks of the neurons of interest in two orthogonal, horizontal, and frontal orientations. Using the standard techniques, stain dissected fly brains with rabbit anti-GFP and an antibody that labels photoreceptor axons primary antibodies. Then label the primary antibodies with fluorescent secondary antibodies and clear the brain in 70%glycerol and 1X PBS.

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