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A novel method for quantifying axon degeneration

by Gibon, Julien , de Léon, Andrés , Hallett, Robin M. , Johnstone, Aaron D. , Carturan, Bruno , Barker, Philip A.

in Animal care / Apoptosis / Automation / Axons / Biology / Biology and Life Sciences / Calcium (extracellular) / Calcium ions / Care and treatment / Cell culture / Cellular signal transduction / Cellular structure / Cytoskeleton / Degeneration / Disease / Dismantling / Dorsal root ganglia / Embryos / Fluorescence microscopy / Ganglia / Growth factors / Health aspects / Internet / Kinases / Measurement methods / Medicine and Health Sciences / Nerve growth factor / Nervous system / Nervous system diseases / Neurodegeneration / Neurodegenerative diseases / Neurological diseases / Neurons / Neuroprotection / Neurosurgery / Pharmacology / Physical Sciences / Physiology / Research and Analysis Methods

2018

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A novel method for quantifying axon degeneration

by Gibon, Julien , de Léon, Andrés , Hallett, Robin M. , Johnstone, Aaron D. , Carturan, Bruno , Barker, Philip A.

2018

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A novel method for quantifying axon degeneration

by Gibon, Julien , de Léon, Andrés , Hallett, Robin M. , Johnstone, Aaron D. , Carturan, Bruno , Barker, Philip A.

2018

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Journal Article

A novel method for quantifying axon degeneration

Gibon, Julien,

de Léon, Andrés,

Hallett, Robin M.,

Johnstone, Aaron D.,

Carturan, Bruno,

Barker, Philip A.

2018

Overview

Axons normally degenerate during development of the mammalian nervous system, but dysregulation of the same genetically-encoded destructive cellular machinery can destroy crucial structures during adult neurodegenerative diseases. Nerve growth factor (NGF) withdrawal from dorsal root ganglia (DRG) axons is a well-established in vitro experimental model for biochemical and cell biological studies of developmental degeneration. Definitive methods for measuring axon degeneration have been lacking and here we report a novel method of axon degeneration quantification from bulk cultures of DRG that enables objective and automated measurement of axonal density over the entire field of radial axon outgrowth from the ganglion. As proof of principal, this new method, written as an R script called Axoquant 2.0, was used to examine the role of extracellular Ca2+ in the execution of cytoskeletal disassembly during degeneration of NGF-deprived DRG axons. This method can be easily applied to examine degenerative or neuroprotective effects of gene manipulations and pharmacological interventions.

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Publisher

Public Library of Science,Public Library of Science (PLoS)

Subject

Animal care

/ Apoptosis

/ Automation

/ Axons

/ Biology

/ Biology and Life Sciences

/ Calcium (extracellular)