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Analysis of Astrocyte Territory Volume and Tiling in Thick Free-Floating Tissue Sections

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Quantifying Filopodia in Cultured Astrocytes by an Algorithm.

Georg Aumann^1,2, Felix Friedländer³, Matthias Thümmler⁴

¹Institute of Anatomy, University of Dresden, Dresden, Germany.

Neurochemical Research

|March 1, 2017

Summary

This summary is machine-generated.

Researchers developed a new automated method, the Filopodia Specific Shape Factor (FSSF), to quantify astrocyte filopodia dynamics. This tool aids in understanding glia-neuronal communication and the molecular basis of fine cellular structures.

Keywords:

Actin cytoskeleton Cell culture Cell motility Glia-synaptic interaction Glutamate Morphometry

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Area of Science:

Neuroscience
Cell Biology
Glial Cell Biology

Background:

Astrocytes extend fine peripheral astrocyte processes (PAPs) around synapses, crucial for glia-neuronal communication.
The stimuli, mechanisms, and molecular basis for PAP formation and dynamics remain unclear.
Investigating these processes in vitro using glial filopodia is essential.

Purpose of the Study:

To introduce and detail an automated method for filopodia morphometry.
To quantify filopodia dynamics and morphology in astrocytes.
To provide a tool for studying glia-neuronal communication and PAP formation.

Main Methods:

Development and detailed description of the Filopodia Specific Shape Factor (FSSF) algorithm for ImageJ.
Automated image processing and measurement of filopodia number and length.
Validation of FSSF, including influence of thresholding and pixel grid; analysis of substance-induced filopodia dynamics.

Main Results:

The FSSF quantifies filopodia independently of overall cell shape or size.
Filopodia formation is sensitive to various stimuli (glutamate, mGluR agonists, EGF) and environmental factors (pH, culture duration).
Exemplary results demonstrate substance-induced filopodia dynamics.

Conclusions:

The FSSF is a robust tool for automated measurement of astrocyte filopodia.
This method facilitates the study of mechanisms underlying PAP formation and glia-neuronal communication.
The FSSF has potential applications beyond astrocytes, including neuronal growth cones and other cell types.