Toward an improved understanding of the role of nicotinic acetylcholine receptors (nAChR) in development and in congenital or degenerative disorders of the mammalian CNS, the following studies are proposed. Biochemical studies will be continued on sites in mature rat brain subcellular fractions that interact with putative nAChR-specific probes. These studies shall involve the use of curaremimetic neurotoxins, antibodies raised against nAChR from the electric organ of ray or eel, and affinity reagents that specifically label nAChR in muscle and electric tissue. These probes for nAChR will be used in established ligand binding assays, affinity labeling protocols and affinity purification procedures.
The aim of these studies is to elucidate the molecular features of and interrelationships between binding sites for putative nAChR probes in the mammalian CNS. Continued maintenance of the human medulloblastoma clonal line, TE671, in cell culture is proposed. Levels of expression by cultured TE671 cells of toxin and affinity reagent binding sites and nAChR-like antigenic determinants will be quantitated and compared by use of established radioligand binding and immunoassays. The objectives of these studies are to assess the relative density of ligand and antibody binding sites on the TE671 continuous line. Molecular features of sites that are affinity purified or affinity labeled by interaction with putative nAChR probes will be elucidated. Isotopic ion efflux studies have been established and will continue to be used to detect the functional response of the ensemble of physiologically-relevant nAChR on TE671 cells. Detailed investigation of small ligand, toxin and antibody actions on nAChR functional responses will be undertaken toward elucidation of the relationship between functional nAChR and binding sites for putative receptor probes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS016821-06
Application #
3397156
Study Section
Neurological Sciences Subcommittee 1 (NLS)
Project Start
1981-09-01
Project End
1990-06-30
Budget Start
1988-07-01
Budget End
1990-06-30
Support Year
6
Fiscal Year
1988
Total Cost
Indirect Cost
Name
St. Joseph's Hosp/Medical Center (Phoenix)
Department
Type
DUNS #
City
Phoenix
State
AZ
Country
United States
Zip Code
85013
Lukas, R J (1995) Diversity and patterns of regulation of nicotinic receptor subtypes. Ann N Y Acad Sci 757:153-68
Lukas, R J (1993) Expression of ganglia-type nicotinic acetylcholine receptors and nicotinic ligand binding sites by cells of the IMR-32 human neuroblastoma clonal line. J Pharmacol Exp Ther 265:294-302
Joy, A M; Siegel, H N; Lukas, R J (1993) Photoaffinity labeling of muscle-type nicotinic acetylcholine receptors and neuronal/nicotinic alpha-bungarotoxin binding sites with a derivative of alpha-bungarotoxin. Brain Res Mol Brain Res 17:95-100
Lukas, R J; Bencherif, M (1992) Heterogeneity and regulation of nicotinic acetylcholine receptors. Int Rev Neurobiol 34:25-131
Lukas, R J (1991) Effects of chronic nicotinic ligand exposure on functional activity of nicotinic acetylcholine receptors expressed by cells of the PC12 rat pheochromocytoma or the TE671/RD human clonal line. J Neurochem 56:1134-45
Bencherif, M; Lukas, R J (1991) Ligand binding and functional characterization of muscarinic acetylcholine receptors on the TE671/RD human cell line. J Pharmacol Exp Ther 257:946-53
Lukas, R J (1990) Heterogeneity of high-affinity nicotinic [3H]acetylcholine binding sites. J Pharmacol Exp Ther 253:51-7
Lukas, R J; Audhya, T; Goldstein, G et al. (1990) Interactions of the thymic polypeptide hormone thymopoietin with neuronal nicotinic alpha-bungarotoxin binding sites and with muscle-type, but not ganglia-type, nicotinic acetylcholine receptor ligand-gated ion channels. Mol Pharmacol 38:887-94
Oswald, R E; Papke, R L; Lukas, R J (1989) Characterization of nicotinic acetylcholine receptor channels of the TE671 human medulloblastoma clonal line. Neurosci Lett 96:207-12
Lukas, R J (1989) Pharmacological distinctions between functional nicotinic acetylcholine receptors on the PC12 rat pheochromocytoma and the TE671 human medulloblastoma. J Pharmacol Exp Ther 251:175-82

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