This is a continuation of a long term study of the nerve growth factor protein, NFG its binding to its receptor, NGFR, and NGF effects relevant to regeneration. NGF plays a role in the development, maintenance and regeneration of some PNS and CNS neurons. NGF has ameliorative effects on certain lesioned neural structures where it plays a role in axon sprouting and regeneration. The primary event associated with NGF action is binding to NGFR. Subsequently, there is a broad spectrum of NGF effects on target tissues. Two hypotheses are to be tested. One is that NGFR from different species or tissues is made up of similar proteins but different carbohydrate compositions that affect NGF binding. The other hypothesis is that NGF has a regulatory role on oxidant-antioxidant metabolism that is relevant to regenerative outcome. The initial binding of NGF to NGFR in CNS tissues is to be compared to that already established for PNS and the NGF-responsive neuronal cell lines, PC12, SY5Y and LAN-1. NGF binding will be characterized using a soluble receptor assay that uses iodinated beta NGF as ligand. Additional ligands to be used as controls will be cytochrome C, insulin and EGF as well as synthetic peptides homologous to part of the NGF sequence. NGFR will be immunoprecipitated using two monoclonal antibodies, the MC192 to rodent NGFR, and a well established monoclonal antibody to human NGFR. Immunoprecipitated NGFR will be characterized by SDS-PAGE, preparative electrofocusing and HPLC. NGFR apoprotein and sugar moieties will be perturbed with retinoic acid, tunicamycin and lectins to test effects on receptor activity and structure. NGF induces the antioxidant enzymes glutathione peroxidase and catalase in vitro. Thus, the mechanism of NGF sparing of lesioned cells, due to regulatory effects on oxidant-antioxidant metabolism, will continue to be studied in the NGF sensitive cell lines PC12 and SY5Y. ln rats, spinal hemisection, or a regimen of exposure to anti-NGF has irreversible developmental consequences on sensory neurons. There is a decrease in the number of neurons and an increase in sensory axons. Here, antibodies to and assays for key antioxidants will be applied to both the in vitro and in vivo paradigms. Because of its effects on CNS cholinergic neurons involved in Alzheimers, it has been suggested that expression of NGF activity may be one factor in the development of this disease. It has also been suggested that an understanding of NGF action may be relevant to aspects of neurofibromatosis, ALS and CNS trauma.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
2R01NS018708-07
Application #
3398732
Study Section
Neurology B Subcommittee 1 (NEUB)
Project Start
1982-07-01
Project End
1991-08-31
Budget Start
1988-09-01
Budget End
1989-08-31
Support Year
7
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Texas Medical Br Galveston
Department
Type
Schools of Medicine
DUNS #
041367053
City
Galveston
State
TX
Country
United States
Zip Code
77555
Turner, C P; Perez-Polo, J R (1998) Expression of the low affinity neurotrophin receptor, P75NGFR, in the rat forebrain, following unilateral bulbectomy. Int J Dev Neurosci 16:527-38
Toliver-Kinsky, T; Papaconstantinou, J; Perez-Polo, J R (1997) Age-associated alterations in hippocampal and basal forebrain nuclear factor kappa B activity. J Neurosci Res 48:580-7
Tong, L; Werrbach-Perez, K; Perez-Polo, J R (1997) Retinoic acid induces apoptosis in PC12 cells independent of neurotrophic factors. J Neurochem 68:1424-35
Sampath, D; Perez-Polo, R (1997) Regulation of antioxidant enzyme expression by NGF. Neurochem Res 22:351-62
Gottesfeld, Z; Simpson, S; Yuwiler, A et al. (1996) Effects of nerve growth factor on splenic norepinephrine and pineal N-acetyl-transferase in neonate rats exposed to alcohol in utero: neuroimmune correlates. Int J Dev Neurosci 14:655-62
Yu, J; Wiley, R G; Perez-Polo, R J (1996) Altered NGF protein levels in different brain areas after immunolesion. J Neurosci Res 43:213-23
Pan, Z; Perez-Polo, R (1996) Regulation of gamma-glutamylcysteine synthetase activity by nerve growth factor. Int J Dev Neurosci 14:559-66
Pan, Z; Perez-Polo, R (1996) Increased uptake of L-cysteine and L-cystine by nerve growth factor in rat pheochromocytoma cells. Brain Res 740:21-6
Taglialatela, G; Hibbert, C J; Hutton, L A et al. (1996) Suppression of p140trkA does not abolish nerve growth factor-mediated rescue of serum-free PC12 cells. J Neurochem 66:1826-35
Tong, L; Perez-Polo, J R (1996) Effect of nerve growth factor on AP-1, NF-kappa B, and Oct DNA binding activity in apoptotic PC12 cells: extrinsic and intrinsic elements. J Neurosci Res 45:1-12

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