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Journal of Neuroscience, Vol 9, 3505-3512, Copyright © 1989 by Society for Neuroscience
Expression of growth-associated protein B-50 (GAP43) in dorsal root ganglia and sciatic nerve during regenerative sprouting
CE Van der Zee, HB Nielander, JP Vos, S Lopes da Silva, J Verhaagen, AB Oestreicher, LH Schrama, P Schotman and WH Gispen
Division of Molecular Neurobiology, Rudolf Magnus Institute, University of Utrecht, The Netherlands.
Recently it has been shown that B-50 is identical to the neuron- specific,
growth-associated protein GAP43. The present study reports on the fate of
B-50/GAP43 mRNA and B-50/GAP43 protein, determined by radioimmunoassay, in
a rat model of peripheral nerve regeneration (sciatic nerve crush) over a
period of 37 and 312 d, respectively. Moreover, the effects of repeated
subcutaneous injection of the neurotrophic peptide Org.2766 (an ACTH4-9
analog) and of a conditioning lesion on B-50/GAP43 protein levels in the
regenerating nerve and dorsal root ganglia (DRG) were investigated. Both
treatments enhanced the functional recovery as evidenced by a foot-flick
withdrawal test. Immunocytochemical analysis using antineurofilament
antibodies revealed a peptide-induced increase in the number of outgrowing
sprouts in the sciatic nerve. Both the peptide and the conditioning lesion
amplified the crush lesion-induced increase in B-50 protein content in the
nerve as determined by radioimmunoassay. B-50 protein levels seem to
correlate proportionally with the number of sprouts. In the DRG of the
crushed sciatic nerve, the time course of B-50 expression was studied. B-50
mRNA was quantified from Northern blots. A linear increase up to 10 times
the basal level of B-50 mRNA was observed 2 d postsurgery, followed by a
gradual decline to normal levels at day 37. The first significant rise in
B-50 mRNA level became apparent between 8 and 16 hr after placement of the
crush lesion. The first significant rise in B-50 protein level occurred 40
hr after the crush lesion, reaching a plateau of 3 times the basal level
between day 6 and 20. B-50 protein levels in DRG cell bodies remained
elevated up to 60 d after crush, a period much longer than that observed
for B-50 mRNA. Thus, during a later phase of peripheral axonal
regeneration, the presence of B-50 appears to be prolonged, probably by an
increase in half-life and not so much by enhanced transcription. Treatment
with Org.2766 did not affect the B- 50/GAP43 levels in DRG cell bodies
during the first 6 d following crush. Conditioning lesion resulted in a DRG
B-50/GAP43 protein amount at the same level as in rats 14 d after the test
lesion. B-50/GAP43 levels in DRG are probably influenced by the rapid
axonal transport of the protein, as has been reported by others.
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