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Use of explant cultures of peripheral nerves of adult vertebrates to study axonal regeneration in vitro

Tonge, David; Edström, Anders LU and Ekström, Per LU (1998) In Progress in Neurobiology 54(4). p.459-480
Abstract

Explanted preparations of peripheral nerves with attached dorsal root ganglia of adult mammals and amphibia survive for several days in serum-free medium and can be used to study axonal regeneration in vitro. This review outlines the methods which we routinely use and how they may be applied to study different aspects of axonal regeneration. When the peripheral nerves are crushed in vitro, axons regenerate through the crush site into the distal stump within 1 day (mouse) or 3 days (frog). The outgrowth distance of the leading sensory axons can be determined with the use of a simple method based on axonal transport of labelled proteins. A compartmentalised system permits selective application of drugs and other agents to either ganglia... (More)

Explanted preparations of peripheral nerves with attached dorsal root ganglia of adult mammals and amphibia survive for several days in serum-free medium and can be used to study axonal regeneration in vitro. This review outlines the methods which we routinely use and how they may be applied to study different aspects of axonal regeneration. When the peripheral nerves are crushed in vitro, axons regenerate through the crush site into the distal stump within 1 day (mouse) or 3 days (frog). The outgrowth distance of the leading sensory axons can be determined with the use of a simple method based on axonal transport of labelled proteins. A compartmentalised system permits selective application of drugs and other agents to either ganglia or peripheral nerve containing the regenerating axons and has been used to study selected aspects of regeneration including influence of non-neuronal cells, retrograde signalling, axonal release of proteins during regeneration and the role of phospholipase A2 activity. Explanted preparations may also be cultured in a layer of extracellular matrix material (matrigel), in which spontaneous outgrowth of a large number of naked axons from the cut ends of nerves starts within 1 day and continues for several days. This provides an opportunity to study the direct effects of different agents on axonal elongation. Preparations cultured in collagen gels show sparse spontaneous axonal growth, but this can be increased by addition of certain growth factors. The phenotype of the regenerating axons can be studied using immunohistochemical methods.

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author
organization
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type
Contribution to journal
publication status
published
subject
in
Progress in Neurobiology
volume
54
issue
4
pages
22 pages
publisher
Elsevier
external identifiers
  • Scopus:0032473444
ISSN
0301-0082
DOI
10.1016/S0301-0082(97)00072-5
language
English
LU publication?
yes
id
1e606a2a-0020-4913-8ffc-872bd884ddc4
date added to LUP
2016-12-07 14:43:27
date last changed
2017-02-05 04:55:55
@article{1e606a2a-0020-4913-8ffc-872bd884ddc4,
  abstract     = {<p>Explanted preparations of peripheral nerves with attached dorsal root ganglia of adult mammals and amphibia survive for several days in serum-free medium and can be used to study axonal regeneration in vitro. This review outlines the methods which we routinely use and how they may be applied to study different aspects of axonal regeneration. When the peripheral nerves are crushed in vitro, axons regenerate through the crush site into the distal stump within 1 day (mouse) or 3 days (frog). The outgrowth distance of the leading sensory axons can be determined with the use of a simple method based on axonal transport of labelled proteins. A compartmentalised system permits selective application of drugs and other agents to either ganglia or peripheral nerve containing the regenerating axons and has been used to study selected aspects of regeneration including influence of non-neuronal cells, retrograde signalling, axonal release of proteins during regeneration and the role of phospholipase A<sub>2</sub> activity. Explanted preparations may also be cultured in a layer of extracellular matrix material (matrigel), in which spontaneous outgrowth of a large number of naked axons from the cut ends of nerves starts within 1 day and continues for several days. This provides an opportunity to study the direct effects of different agents on axonal elongation. Preparations cultured in collagen gels show sparse spontaneous axonal growth, but this can be increased by addition of certain growth factors. The phenotype of the regenerating axons can be studied using immunohistochemical methods.</p>},
  author       = {Tonge, David and Edström, Anders and Ekström, Per},
  issn         = {0301-0082},
  language     = {eng},
  month        = {03},
  number       = {4},
  pages        = {459--480},
  publisher    = {Elsevier},
  series       = {Progress in Neurobiology},
  title        = {Use of explant cultures of peripheral nerves of adult vertebrates to study axonal regeneration in vitro},
  url          = {http://dx.doi.org/10.1016/S0301-0082(97)00072-5},
  volume       = {54},
  year         = {1998},
}