A novel method for quantifying axon degeneration

PloS One
Aaron D JohnstonePhilip A Barker

Abstract

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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Citations

Nov 4, 2020·Journal of Applied Toxicology : JAT·Anshul Srivastava, Joseph P Hanig
Aug 24, 2020·Neuroscience Letters·Franziska Gross, Nurcan Üçeyler

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Methods Mentioned

BETA
gene
chemical treatments

Software Mentioned

NIH ImageJ
Studio
GraphPad Prism
Zeiss
R Studio
Axoquant
R script
R
Zen

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