This grant proposes a series of related studies on the synthesis and processing of the neuronal trophic factor Nerve Growth Factor (NGF) in sites that are important for neuronal development. Using a cDNA probe encoding the mouse beta-NGF gene, studies will be continued on identifying tissues that synthesize NGF in the adult and during development. We will attempt to identify the cell types that synthesize NGF within these tissues by examining NGF gene expression in defined cell types in culture and by doing in situ hybridization on tissue sections using an NGF gene probe. The NGF probe will be used to quantify NGF mRNA levels during the development of tissues innervated by sympathetic and sensory neurons. We will determine whether there are relationships between the synthesis of NGF mRNA and the induction of sumpathetic neurons, the development and extent of innervation in targets, and neuronal cell death. We will determine in adult tissues whether denervation, already known to increase NGF levels, does so by increasing beta-NGF gene expression. In a second set of experiments, we will examine the form of NGF synthesized in tissues and cells that express the gene using antibodies that are specific for amino acid segments in the precursor. Residues between these segments are cleaved during normal processing in exocrine cells in the male mouse submaxillary gland. We will see whether the same processing steps occur in the tissues and cells believed to be the physiological sources of NGF. Some of these cells are known not to process other prohormones, so the precursor may be important. It will be purified and its properties will be examined. If NGF is processed in physiological sources, we will determine whether proteases are able to process the prohormone in vitro are present at appropriate sites in vivo. Antibodies specific for amino acid residues in the precursor will be used to purify the three polypeptides produced in addition to NGF by processing of the prohormone in the mouse submaxillary gland. These purified polypeptides will be tested for biological actions on NGF-responsive cells and will be used to make antibodies for studies on processing of proNGF. These results should provide information on how NGF, an important trophic hormone, controls the development of responsive neuronal cell populations.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS021824-02
Application #
3403460
Study Section
Neurological Sciences Subcommittee 1 (NLS)
Project Start
1984-12-01
Project End
1987-11-30
Budget Start
1985-12-01
Budget End
1986-11-30
Support Year
2
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of California San Francisco
Department
Type
Schools of Medicine
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Barker, P A; Miller, F D; Large, T H et al. (1991) Generation of the truncated form of the nerve growth factor receptor by rat Schwann cells. Evidence for post-translational processing. J Biol Chem 266:19113-9
Bossy, B; Bossy-Wetzel, E; Reichardt, L F (1991) Characterization of the integrin alpha 8 subunit: a new integrin beta 1-associated subunit, which is prominently expressed on axons and on cells in contact with basal laminae in chick embryos. EMBO J 10:2375-85
Weskamp, G; Reichardt, L F (1991) Evidence that biological activity of NGF is mediated through a novel subclass of high affinity receptors. Neuron 6:649-63
Bossy, B; Reichardt, L F (1990) Chick integrin alpha V subunit molecular analysis reveals high conservation of structural domains and association with multiple beta subunits in embryo fibroblasts. Biochemistry 29:10191-8
Roback, J D; Large, T H; Otten, U et al. (1990) Nerve growth factor expression in the developing hippocampus isolated in vitro. Dev Biol 137:451-5
Large, T H; Weskamp, G; Helder, J C et al. (1989) Structure and developmental expression of the nerve growth factor receptor in the chicken central nervous system. Neuron 2:1123-34
Clegg, D O; Large, T H; Bodary, S C et al. (1989) Regulation of nerve growth factor mRNA levels in developing rat heart ventricle is not altered by sympathectomy. Dev Biol 134:30-7
Reichardt, L F; Bixby, J L; Hall, D E et al. (1989) Integrins and cell adhesion molecules: neuronal receptors that regulate axon growth on extracellular matrices and cell surfaces. Dev Neurosci 11:332-47
Shelton, D L; Reichardt, L F (1986) Studies on the expression of the beta nerve growth factor (NGF) gene in the central nervous system: level and regional distribution of NGF mRNA suggest that NGF functions as a trophic factor for several distinct populations of neurons. Proc Natl Acad Sci U S A 83:2714-8
Shelton, D L; Reichardt, L F (1986) Studies on the regulation of beta-nerve growth factor gene expression in the rat iris: the level of mRNA-encoding nerve growth factor is increased in irises placed in explant cultures in vitro, but not in irises deprived of sensory or sympathetic innervat J Cell Biol 102:1940-8

Showing the most recent 10 out of 11 publications