Central facts about nerve growth factor (NGF) have yet to be established. Despite numerous studies elucidating the protein's unique ability to promote and maintain peripheral nerve function, the site of NGF's synthesis within the body remains unknown. Cells from many different species and organs synthesize and secrete NGF-like molecules in culture, giving rise to the theory that all peripheral tissues produce the protein in vivo. The first goal of our work will be to investigate that hypothesis. Selected tissues in the mouse which are innervated by sympathetic, sensory, and motor neurons will be denervated in order to stimulate NGF production, and radioimmunoassays and bioassay will measure levels of the protein. NGF-containing cells will be identified by light and electron microscopy using antibodies to NGF coupled with fluorescent dyes and colloidal gold. Secondly, we seek to establish whether peripheral tissues in vivo and mouse cells in culture produce similar forms of NGF and whether these proteins are identical to the well-characterized NGF in mouse salivary glands. Tissue extracts and conditioned medium will be electrophoresed on polyacrylamide gels in dissociating and non-dissociating solvents. Cells will be analyzed by the """"""""western blot"""""""" technique, utilizing as probes antibodies raised against the Alpha, Beta and Gamma subunits of the 7S NGF complex from submandibular glands. Results from these experiments should provide a more complete understanding of NGF's distribution in peripheral tissues and its molecular properties. These data will fill significant gaps in our knowledge as we move towards an ultimate understanding of NGF's role in nerve development, function, and repair.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
3R01NS019100-03S1
Application #
3399111
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1983-01-01
Project End
1986-12-31
Budget Start
1986-01-01
Budget End
1986-12-31
Support Year
3
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Harvard University
Department
Type
Schools of Medicine
DUNS #
082359691
City
Boston
State
MA
Country
United States
Zip Code
02115
Murphy, R A; Chlumecky, V; Smillie, L B et al. (1989) Isolation and characterization of a glycosylated form of beta nerve growth factor in mouse submandibular glands. J Biol Chem 264:12502-9
Siminoski, K; Murphy, R A; Rennert, P et al. (1987) Cortisone, testosterone, and aldosterone reduce levels of nerve growth factor messenger ribonucleic acid in L-929 fibroblasts. Endocrinology 121:1432-7
Gonnella, P A; Siminoski, K; Murphy, R A et al. (1987) Transepithelial transport of epidermal growth factor by absorptive cells of suckling rat ileum. J Clin Invest 80:22-32
Murphy, R A; Landis, S C; Bernanke, J et al. (1986) Absence of the alpha and gamma subunits of 7S nerve growth factor in denervated rodent iris: immunocytochemical studies. Dev Biol 114:369-80
Siminoski, K; Gonnella, P; Bernanke, J et al. (1986) Uptake and transepithelial transport of nerve growth factor in suckling rat ileum. J Cell Biol 103:1979-90
Siminoski, K; Bernanke, J; Kay, C et al. (1986) Steroids and triiodothyronine reduce nerve growth factor concentrations in medium conditioned by L-929 fibroblasts. Endocrinology 118:1417-25
Watson, A Y; Anderson, J K; Siminoski, K et al. (1985) Cellular and subcellular colocalization of nerve growth factor and epidermal growth factor in mouse submandibular glands. Anat Rec 213:365-76
Finke, U; Rutten, M; Murphy, R A et al. (1985) Effects of epidermal growth factor on acid secretion from guinea pig gastric mucosa: in vitro analysis. Gastroenterology 88:1175-82