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Involvement of axonal phospholipase A2 activity in the outgrowth of adult mouse sensory axons in vitro

Hornfelt, M. LU ; Ekström, Per LU and Edström, A. LU (1999) In Neuroscience 91(4). p.1539-1547
Abstract

The effect on axonal outgrowth of inhibition of phospholipase A2 activity was studied in a recently developed in vitro model, where dorsal root ganglia with attached spinal roots and nerve stumps from young adult mice were cultured in an extracellular matrix material (Matrigel). The phospholipase A2 inhibitors 4-bromophenacyl bromide and oleyloxyethyl phosphorylcholine dose-dependently reduced axonal outgrowth from the sciatic nerve stump. A similar inhibitory effect was seen when only the cut nerve end was exposed to the inhibitors in a compartmental culture system. The local effect of phospholipase A2 inhibition was further investigated on axons established in culture, using time-lapse recording.... (More)

The effect on axonal outgrowth of inhibition of phospholipase A2 activity was studied in a recently developed in vitro model, where dorsal root ganglia with attached spinal roots and nerve stumps from young adult mice were cultured in an extracellular matrix material (Matrigel). The phospholipase A2 inhibitors 4-bromophenacyl bromide and oleyloxyethyl phosphorylcholine dose-dependently reduced axonal outgrowth from the sciatic nerve stump. A similar inhibitory effect was seen when only the cut nerve end was exposed to the inhibitors in a compartmental culture system. The local effect of phospholipase A2 inhibition was further investigated on axons established in culture, using time-lapse recording. Exposure to phospholipase A2 inhibitors caused the retraction of filopodia extensions and a reduction in growth cone motility within a few minutes. After removal of inhibition, normal growth cone motility and axonal growth were regained. Nerve cell bodies and axons, in contrast to Schwann cells, showed immunoreactivity after staining with an antiserum against secretory phospholipase A2, and elevated levels of the enzyme could be detected after culture for 24 h. The immunoreactive protein was of approximately 170,000 molecular weight (phospholipase A2-170) as determined by sodium dodecyl sulphate- polyacrylamide gel electrophoresis and immunoblotting. The localization of phospholipase A2-170 in axons growing into the Matrigel was also demonstrated by use of a whole-mount technique. The results of this study show the importance of continuous phospholipase A2 activity for growth cone motility and axonal outgrowth in the mammalian peripheral nerve, and suggest the involvement of an axonally localized enzyme.

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author
organization
publishing date
type
Contribution to journal
publication status
published
keywords
Growth cone, Neurite outgrowth, Peripheral nerve, Phospholipase A-170
in
Neuroscience
volume
91
issue
4
pages
1539 - 1547
publisher
Elsevier
external identifiers
  • Scopus:0032905046
ISSN
0306-4522
DOI
10.1016/S0306-4522(98)00684-8
language
English
LU publication?
yes
id
eb720080-137b-4e63-88c6-3e46c6725d12
date added to LUP
2016-12-07 14:45:28
date last changed
2017-01-20 16:29:30
@article{eb720080-137b-4e63-88c6-3e46c6725d12,
  abstract     = {<p>The effect on axonal outgrowth of inhibition of phospholipase A<sub>2</sub> activity was studied in a recently developed in vitro model, where dorsal root ganglia with attached spinal roots and nerve stumps from young adult mice were cultured in an extracellular matrix material (Matrigel). The phospholipase A<sub>2</sub> inhibitors 4-bromophenacyl bromide and oleyloxyethyl phosphorylcholine dose-dependently reduced axonal outgrowth from the sciatic nerve stump. A similar inhibitory effect was seen when only the cut nerve end was exposed to the inhibitors in a compartmental culture system. The local effect of phospholipase A<sub>2</sub> inhibition was further investigated on axons established in culture, using time-lapse recording. Exposure to phospholipase A<sub>2</sub> inhibitors caused the retraction of filopodia extensions and a reduction in growth cone motility within a few minutes. After removal of inhibition, normal growth cone motility and axonal growth were regained. Nerve cell bodies and axons, in contrast to Schwann cells, showed immunoreactivity after staining with an antiserum against secretory phospholipase A<sub>2</sub>, and elevated levels of the enzyme could be detected after culture for 24 h. The immunoreactive protein was of approximately 170,000 molecular weight (phospholipase A<sub>2</sub>-170) as determined by sodium dodecyl sulphate- polyacrylamide gel electrophoresis and immunoblotting. The localization of phospholipase A<sub>2</sub>-170 in axons growing into the Matrigel was also demonstrated by use of a whole-mount technique. The results of this study show the importance of continuous phospholipase A<sub>2</sub> activity for growth cone motility and axonal outgrowth in the mammalian peripheral nerve, and suggest the involvement of an axonally localized enzyme.</p>},
  author       = {Hornfelt, M. and Ekström, Per and Edström, A.},
  issn         = {0306-4522},
  keyword      = {Growth cone,Neurite outgrowth,Peripheral nerve,Phospholipase A-170},
  language     = {eng},
  number       = {4},
  pages        = {1539--1547},
  publisher    = {Elsevier},
  series       = {Neuroscience},
  title        = {Involvement of axonal phospholipase A2 activity in the outgrowth of adult mouse sensory axons in vitro},
  url          = {http://dx.doi.org/10.1016/S0306-4522(98)00684-8},
  volume       = {91},
  year         = {1999},
}