The purpose of this project is to further our understanding of the mechanism by which specific neuronotrophic factors act on the mammalian nervous system during development. To date, this project has dealt exclusively with one neuronotrophic factor, the nerve growth factor (NGF), known to be necessary for the development of vertebrate sympathetic and sensory neurons. NGF is a multimeric protein made up of three dissimilar subunits, only one of which is biologically active. A large number of studies would suggest that there are alterations in ambient levels of NGF in humans suffering from a number of neuropathies including the peripheral and central nervous system. However, there are inconsistencies in some of these reports, difficulty in repeating some of the observations, as well as disagreement as to their significance. This is due to our ignorance of the regulatory mechanisms responsible for NGF synthesis, delivery and mode of action on target cells. These ambiguities also stem from the use of murine-NGF as an antigen and standard in assays on human tissues and the use of a rat non-neuronal adrenal clonal cell line (PC12) or of heterogeneous cell populations in primary cultures in the study of NGF effects. In order to circumvent some of these problems, we will focus our effects on NGF of human origin and NGF-responsiveness neuroblastoma clonal lines of human sympathetic origin. We therefore propose to: (1) To use NGF isolated from human term placenta and monoclonal antibodies developed by us and directed against the beta subunit of human term placental NGF in the conclusive phase of our determination of the quaternary structure of human NGF and its synthesis and intracellular processing. (2) To further characterize the cell surface receptors to NGF using isolation procedures developed by us. (3) To use in vitro differentiating agents, such as retinoic acid and dibutyryl cyclic AMP: and metabolic inhibitors, such as tunicamycin, in order to manipulate expression of NGF receptors in vitro. (4) As part of these studies, to develop monoclonal antibodies directed against receptors to NGF. (5) Lastly, to determine the effects of NGF on neuronal injury in vitro. In particular, we wish to determine the mechanism of the NGF stimulation of neuronal survival following free radical damage.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
2R01NS018708-04
Application #
3398730
Study Section
Biochemistry Study Section (BIO)
Project Start
1982-07-01
Project End
1988-08-31
Budget Start
1985-09-30
Budget End
1986-08-31
Support Year
4
Fiscal Year
1985
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
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
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

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