Recombinant DNA clones corresponding to mRNA's induced during successful axon regeneration will be used to evaluate the hypothesis that axon regeneration in many mammalian CNS neurons is limited by failure to induce expression of certain """"""""growth-associated"""""""" genes. Clones representing putative """"""""growth-associated"""""""" genes will be selected by screening a library of clones - containing cDNA complementary to all the mRNA's translated in regenerating toad retinal ganglion cells - for clones which hybridize more readily with radiolabeled cDNA from regenerating retinal ganglion cells (representing total polysomal mRNA) than with the corresponding cDNA from the normal control neurons. Clones selected in this way will be used as probes for the presence of homologous mRNA's in a broad phylogenetic spectrum of neurons during developmental axon growth, successful regeneration, or abortive regeneration. The hypothesis being tested predicts that all neurons undergoing developmental axon growth or successful axon regeneration will show greatly increased abilities to hybridize with the """"""""growth-associated"""""""" cDNA probes (because of increased amounts of """"""""growth-associated"""""""" mRNA in those cells) compared with normal adult neurons, but that mammalian CNS neurons which do not regenerate their axons will not increase their hybridization with the probes after axotomy. Hybridizations of pulse-labeled RNA to the """"""""growth-associated"""""""" DNA clones, cell-free translation of mRNA from normal and regenerating neurons, and gel electrophoresis of in vivo labeled retrogradely transported proteins will be used to investiage mechanisms by which axon interruption leads to specific changes in neuronal gene expression. The proposed research establishes a genetic definition of the differences between neurons which grow axons successfully and those which fail to regenerate injured axons.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS020178-02
Application #
3400389
Study Section
Neurological Sciences Subcommittee 1 (NLS)
Project Start
1983-12-01
Project End
1986-11-30
Budget Start
1984-12-01
Budget End
1985-11-30
Support Year
2
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Stanford University
Department
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
Smith, D S; Skene, J H (1997) A transcription-dependent switch controls competence of adult neurons for distinct modes of axon growth. J Neurosci 17:646-58
Patterson, S I; Skene, J H (1994) Novel inhibitory action of tunicamycin homologues suggests a role for dynamic protein fatty acylation in growth cone-mediated neurite extension. J Cell Biol 124:521-36
Hess, D T; Slater, T M; Wilson, M C et al. (1992) The 25 kDa synaptosomal-associated protein SNAP-25 is the major methionine-rich polypeptide in rapid axonal transport and a major substrate for palmitoylation in adult CNS. J Neurosci 12:4634-41
Schreyer, D J; Skene, J H (1991) Fate of GAP-43 in ascending spinal axons of DRG neurons after peripheral nerve injury: delayed accumulation and correlation with regenerative potential. J Neurosci 11:3738-51
Goslin, K; Schreyer, D J; Skene, J H et al. (1990) Changes in the distribution of GAP-43 during the development of neuronal polarity. J Neurosci 10:588-602
Skene, J H; Virag, I (1989) Posttranslational membrane attachment and dynamic fatty acylation of a neuronal growth cone protein, GAP-43. J Cell Biol 108:613-24
Skene, J H (1989) Axonal growth-associated proteins. Annu Rev Neurosci 12:127-56
LaBate, M E; Skene, J H (1989) Selective conservation of GAP-43 structure in vertebrate evolution. Neuron 3:299-310
Basi, G S; Jacobson, R D; Virag, I et al. (1987) Primary structure and transcriptional regulation of GAP-43, a protein associated with nerve growth. Cell 49:785-91
Ignatius, M J; Gebicke-Harter, P J; Skene, J H et al. (1986) Expression of apolipoprotein E during nerve degeneration and regeneration. Proc Natl Acad Sci U S A 83:1125-9

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