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Journal of Neuroscience, Vol 9, 1303-1313, Copyright © 1989 by Society for Neuroscience
Axonal transport and localization of B-50/GAP-43-like immunoreactivity in regenerating sciatic and facial nerves of the rat
W Tetzlaff, H Zwiers, K Lederis, L Cassar and MA Bisby
Department of Medical Physiology, University of Calgary, Alberta, Canada.
Neurons that can regenerate their axons following axotomy increase their
synthesis and axonal transport of a growth-associated protein, called
GAP-43, which has been shown to be identical to the synaptic phosphoprotein
B-50. The function of B-50/GAP-43 to the process of regeneration is
unknown. We used a polyclonal, affinity-purified antibody against B-50 to
study the axonal transport and localization of B-50/GAP-43-like
immunoreactivity (B50LI) in the regenerating sciatic and facial nerves of
adult rats. Quantitative data were obtained by densitometry of the B-50
band in immunoblots of nerve segments, which had been run on
SDS-polyacrylamide gels. In the regenerating sciatic nerve, anterograde
accumulation at a collection ligature was 3.0 times higher than retrograde
accumulation. The mobile fraction of B50LI was only 0.28 of total B50LI and
traveled with a mean anterograde velocity of 5.3 mm/hr. B50LI distribution
in the newly regenerated portion of the nerve revealed maximal B50LI levels
midway between the position of the crush and the fastest-growing axons.
Immunocytochemistry of this portion of the nerve demonstrated B50LI to be
associated with regenerating axons but also to a large extent with
extra-axonal structures outlining the Schwann cell bands of Bungner. This
zone of B50LI-positive Schwann cell bands was found to extend more distally
in nerves in which regeneration had processed longer, e.g., up to 5 mm
distal to the crush after 3 d and 8 mm after 4 d. Further distal to this
zone, many fine regenerating axonal profiles could be detected with B-50
antibody, but were neurofilament negative. These findings raise the
possibility of an extra-axonal function of B-50/GAP-43, as this protein
might be secreted from regenerating axons and might play a role in
axon-Schwann cell interactions during axonal maturation.
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