A novel method for quantifying axon degeneration

Autor: Bruno Carturan, Philip A. Barker, Julien Gibon, Robin M. Hallett, Andrés de Léon, Aaron D. Johnstone
Rok vydání: 2018
Předmět:
0301 basic medicine
Embryology
Cell
lcsh:Medicine
Fluorescent Antibody Technique
Immunostaining
Degeneration (medical)
Biochemistry
Physical Chemistry
Nerve Fibers
0302 clinical medicine
Tubulin
Animal Cells
Ganglia
Spinal

Nerve Growth Factor
Medicine and Health Sciences
lcsh:Science
Cytoskeleton
Egtazic Acid
Cells
Cultured

Neurons
Staining
Neuronal Death
Multidisciplinary
Cell Death
Chelation
Neurodegenerative Diseases
Immunohistochemistry
Molecular Imaging
Ganglion
Chemistry
medicine.anatomical_structure
Neurology
Cell Processes
Physical Sciences
Cellular Types
Anatomy
Research Article
Biology
Research and Analysis Methods
Neuroprotection
03 medical and health sciences
Tubulins
Extracellular
medicine
Calcium Chelating Agents
Chemical Bonding
lcsh:R
Embryos
Biology and Life Sciences
Proteins
Cell Biology
Axons
Cytoskeletal Proteins
Biological Tissue
030104 developmental biology
Nerve growth factor
nervous system
Specimen Preparation and Treatment
Cellular Neuroscience
lcsh:Q
Calcium
Ganglia
Neuroscience
Biomarkers
030217 neurology & neurosurgery
Developmental Biology
Zdroj: PLoS ONE
PLoS ONE, Vol 13, Iss 7, p e0199570 (2018)
ISSN: 1932-6203
Popis: 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.
Databáze: OpenAIRE