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Journal of Neuroscience, Vol 9, 3766-3775, Copyright © 1989 by Society for Neuroscience
Axotomy-induced changes in the expression of a type III neuronal intermediate filament gene
MM Oblinger, J Wong and LM Parysek
Department of Cell Biology and Anatomy, Chicago Medical School, Illinois 60064.
The effect of axotomy on the expression of the 57 kDa neuronal intermediate
filament (IF) protein in adult rat dorsal root ganglion (DRG) neurons was
examined. This IF protein is known to have an exclusively neuronal
localization but is considerably more limited in its distribution in the
nervous system than the neurofilament (NF) triplet proteins. The 57 kDa
neuronal IF protein is similar (and perhaps identical) to the protein
"peripherin" and is known to be the product of a Type III IF gene. Since
the down-regulated expression of NF proteins (products of type IV IF genes)
has been well established, it was of interest to determine whether the
novel 57 kDa IF protein was regulated in a similar or different manner from
that of the NFs in axotomized neurons. In vitro pulse-labeling of DRGs with
35S- methionine: cysteine followed by 2-dimensional gel
electrophoresis/fluorography revealed that the synthesis of the 57 kDa
neuronal IF protein was increased 2 weeks after sciatic nerve crush.
Immunocytochemical studies using a polyclonal antibody to the 57 kDa
neuronal IF protein showed that the immunodetectable levels of this protein
increased in DRG neurons after peripheral axotomy. In the normal DRG,
staining was localized almost exclusively to small-sized neurons. At 2
weeks after axotomy, however, large- and medium-sized neurons also became
immunoreactive; in addition, the overall level of staining in the DRG was
greater than normal. Quantitative analysis of in situ hybridizations of DRG
neurons with a 35S-labeled cDNA probe specific for the 57 kDa neuronal IF
protein revealed a significant increase in the level of 57 kDa IF mRNA in
the large-sized (greater than 1000 microns2) neurons 2 weeks after axotomy;
the level of 57 kDa IF mRNA in the small neurons was not different from
normal at that time. Finally, using a newly developed paradigm for
examining the composition of regenerating axons by axonal transport, we
determined that significant amounts of the 57 kDa neuronal IF protein were
conveyed into the regrowing axonal sprouts of DRG neurons. When DRG neurons
were conditioned by a previous axotomy (a crush axotomy of the distal
sciatic nerve 2 weeks earlier) and then stimulated to regenerate axons by a
second crush axotomy located very close to the DRG, the regenerating
sprouts incorporated and conveyed significantly more 57 kDa IF protein by
slow axonal transport than did those elaborated by unprimed DRG
neurons.(ABSTRACT TRUNCATED AT 400 WORDS)
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