Abstract

The aim of this project is to uncover the mechanism of action of nerve growth factor (NGF), with particular emphasis on NGF-promoted initiation, locomotion, maintenance and regeneration of neurites. Experiments will principally employ cultured sympathetic neurons and the NGF-responsive PC12 pheochromocytoma line developed in this laboratory. The proposed investigation will draw heavily upon existing dat produced under this grant. The following four major separable, but integrated subsets of experimnal topics shall be pursued. I. The transductional mechanisms of NGF. The means by which binding of nGF to its receptors triggers subsequent actionof the factor will be studied. The roles of phosphorylation, intracellular Ca++ activity and methylation will be explored in particular. II. Short-latency, local transcription-independent actions of NGF. This will include the rapid activation of tyrosine hydroxylase in response to NGF and the underlying role of phosphorylation in this phenomenon. Also in this subset, video time lapse recording, scanning EM and biochemical analyses will be employed to describe and mechanistically probe the local regulation by NGF of growth cone shape, motility and locomotion. III. Delayed, transcription-dependent actions of NGF. The induction of microtubule-associated proteins (MAPs) by NGF will be further investigated as well as the role of these proteins in the mechanism by which NGF promotes neurite outgrowth. IV. Genetic analysis of the NGF mechanism. The PC12 line is especially well suited to such an approach. Mutants fo PC12 cells deficient in specific responses to NGF (especially neurite outgrowth) will be generated, selected and characterized. These will be exploited so as to ultimately uncover the identities and functional roles of each of the gene products involved in NGF's actions. As a whole, the above studies relate to the basic mechanisms by which nerves are stimulated to grow and regenerate and have relevance to a number of issues in cellular neurobiology.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS016036-07
Application #
3396632
Study Section
Neurology B Subcommittee 1 (NEUB)
Project Start
1979-07-01
Project End
1986-06-30
Budget Start
1985-07-01
Budget End
1986-06-30
Support Year
7
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
New York University
Department
Type
Schools of Medicine
DUNS #
004514360
City
New York
State
NY
Country
United States
Zip Code
10012

  Publications

Angelastro, J M; Ryu, E J; Torocsik, B et al. (2002) Blue-white selection step enhances the yield of SAGE concatemers. Biotechniques 32:484, 486
Angelastro, J M; Moon, N Y; Liu, D X et al. (2001) Characterization of a novel isoform of caspase-9 that inhibits apoptosis. J Biol Chem 276:12190-200
Padmanabhan, J; Park, D S; Greene, L A et al. (1999) Role of cell cycle regulatory proteins in cerebellar granule neuron apoptosis. J Neurosci 19:8747-56
Anderson, B L; Boldogh, I; Evangelista, M et al. (1998) The Src homology domain 3 (SH3) of a yeast type I myosin, Myo5p, binds to verprolin and is required for targeting to sites of actin polarization. J Cell Biol 141:1357-70
O'Driscoll, K R; Teng, K K; Fabbro, D et al. (1995) Selective translocation of protein kinase C-delta in PC12 cells during nerve growth factor-induced neuritogenesis. Mol Biol Cell 6:449-58
Volonte, C; Greene, L A (1995) Nerve growth factor-activated protein kinase N modulates the cAMP-dependent protein kinase. J Neurosci Res 40:108-16
Ip, N Y; Stitt, T N; Tapley, P et al. (1993) Similarities and differences in the way neurotrophins interact with the Trk receptors in neuronal and nonneuronal cells. Neuron 10:137-49
Volonte, C; Greene, L A (1992) 6-Methylmercaptopurine riboside is a potent and selective inhibitor of nerve growth factor-activated protein kinase N. J Neurochem 58:700-8
Loeb, D M; Tsao, H; Cobb, M H et al. (1992) NGF and other growth factors induce an association between ERK1 and the NGF receptor, gp140prototrk. Neuron 9:1053-65
Volonte, C; Greene, L A (1992) Nerve growth factor-activated protein kinase N. Characterization and rapid near homogeneity purification by nucleotide affinity-exchange chromatography. J Biol Chem 267:21663-70

Showing the most recent 10 out of 46 publications