PUBLICATION

Motor Nerve Transection and Time-lapse Imaging of Glial Cell Behaviors in Live Zebrafish

Authors
Lewis, G.M., and Kucenas, S.
ID
ZDB-PUB-130722-25
Date
2013
Source
Journal of visualized experiments : JoVE   (76): (Journal)
Registered Authors
Kucenas, Sarah
Keywords
none
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Nerve Regeneration/physiology*
  • Neuroglia/pathology
  • Neuroglia/physiology*
  • Peripheral Nervous System/cytology
  • Peripheral Nervous System/injuries*
  • Peripheral Nervous System/physiology
  • Time-Lapse Imaging/methods*
  • Zebrafish
PubMed
23852039 Full text @ J. Vis. Exp.
Abstract

The nervous system is often described as a hard-wired component of the body even though it is a considerably fluid organ system that reacts to external stimuli in a consistent, stereotyped manner, while maintaining incredible flexibility and plasticity. Unlike the central nervous system (CNS), the peripheral nervous system (PNS) is capable of significant repair, but we have only just begun to understand the cellular and molecular mechanisms that govern this phenomenon. Using zebrafish as a model system, we have the unprecedented opportunity to couple regenerative studies with in vivo imaging and genetic manipulation. Peripheral nerves are composed of axons surrounded by layers of glia and connective tissue. Axons are ensheathed by myelinating or non-myelinating Schwann cells, which are in turn wrapped into a fascicle by a cellular sheath called the perineurium. Following an injury, adult peripheral nerves have the remarkable capacity to remove damaged axonal debris and re-innervate targets. To investigate the roles of all peripheral glia in PNS regeneration, we describe here an axon transection assay that uses a commercially available nitrogen-pumped dye laser to axotomize motor nerves in live transgenic zebrafish. We further describe the methods to couple these experiments to time-lapse imaging of injured and control nerves. This experimental paradigm can be used to not only assess the role that glia play in nerve regeneration, but can also be the platform for elucidating the molecular mechanisms that govern nervous system repair.

Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping