Abstract

The objective of this research program is to determine the characteristics and mechanisms of the regulation of dopamine D-2 receptors. It is crucial that we understand the regulation of D-2 receptors, since idiopathic or drug-induced changes in the density of dopamine receptors are thought to be involved in the pathophysiology or treatment of psychiatric and movement disorders such as schizophrenia, parkinsonism, and tardive dyskinesia. The cloning of a cDNA for the D-2 receptor makes it possible to develop tools that have not been available for the study of D-2 receptor regulation. In particular, cell lines expressing a high density of D-2 receptors are being created by transfecting cells with the cDNA clone, RGB-2.
The aim of this proposal is to develop several cell lines derived from various tissues and to use these cells to determine the mechanisms of desensitization and down-regulation of D-2 receptors. To achieve these goals, the following specific objectives will be met. 1) Four new cell lines expressing D-2 receptors will be created by transfecting cells derived from various tissues with either cDNA or genomic DNA encoding the D-2 receptor. 2) Receptors on the cells will be characterized by radioligand binding, by biochemical assays of adenylate cyclase activity, and by photoaffinity labeling and gel electrophoresis. 3) Conditions for treating cells will be optimized by assessing the possibility of growth-dependent changes in D-2 receptor density. 4) Desensitization of D-2 receptors will be evaluated using four measures of rapid desensitization that assess coupling of receptors to G proteins and subcellular localization of the receptors. 5) The effect on the density of D-2 receptors of treating cells with D-2 receptor agonists will be determined. The time course of agonist-induced loss of receptors will be compared to that of desensitization of adenylate cyclase and to the efficacy of the agonist for inhibition of adenylate cyclase. 6) Agonist-induced changes in levels and transcription of D-2 receptor messenger RNA will be quantified by Northern blot analysis and nuclear runoff experiments.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Project (R01)
Project #
5R01MH045372-03
Application #
3385072
Study Section
Neurosciences Research Review Committee (BPN)
Project Start
1989-09-01
Project End
1993-08-31
Budget Start
1991-09-01
Budget End
1992-08-31
Support Year
3
Fiscal Year
1991
Total Cost
Indirect Cost

  Institution

Name
Oregon Health and Science University
Department
Type
Schools of Dentistry
DUNS #
009584210
City
Portland
State
OR
Country
United States
Zip Code
97239

  Publications

Abraham, Antony D; Neve, Kim A; Lattal, K Matthew (2014) Dopamine and extinction: a convergence of theory with fear and reward circuitry. Neurobiol Learn Mem 108:65-77
Clayton, Cecilea C; Donthamsetti, Prashant; Lambert, Nevin A et al. (2014) Mutation of three residues in the third intracellular loop of the dopamine D2 receptor creates an internalization-defective receptor. J Biol Chem 289:33663-75
Rangel-Barajas, Claudia; Malik, Maninder; Taylor, Michelle et al. (2014) Characterization of [(3) H]LS-3-134, a novel arylamide phenylpiperazine D3 dopamine receptor selective radioligand. J Neurochem 131:418-31
Neve, K A; Ford, C P; Buck, D C et al. (2013) Normalizing dopamine D2 receptor-mediated responses in D2 null mutant mice by virus-mediated receptor restoration: comparing D2L and D2S. Neuroscience 248:479-87
Gantz, Stephanie C; Ford, Christopher P; Neve, Kim A et al. (2011) Loss of Mecp2 in substantia nigra dopamine neurons compromises the nigrostriatal pathway. J Neurosci 31:12629-37
Lan, Hongxiang; Liu, Yong; Bell, Michal I et al. (2009) A dopamine D2 receptor mutant capable of G protein-mediated signaling but deficient in arrestin binding. Mol Pharmacol 75:113-23
Lan, Hongxiang; Teeter, Martha M; Gurevich, Vsevolod V et al. (2009) An intracellular loop 2 amino acid residue determines differential binding of arrestin to the dopamine D2 and D3 receptors. Mol Pharmacol 75:19-26
Liu, Yong; Buck, David C; Neve, Kim A (2008) Novel interaction of the dopamine D2 receptor and the Ca2+ binding protein S100B: role in D2 receptor function. Mol Pharmacol 74:371-8
Liu, Yong; Buck, David C; Macey, Tara A et al. (2007) Evidence that calmodulin binding to the dopamine D2 receptor enhances receptor signaling. J Recept Signal Transduct Res 27:47-65
Watts, Val J; Neve, Kim A (2005) Sensitization of adenylate cyclase by Galpha i/o-coupled receptors. Pharmacol Ther 106:405-21

Showing the most recent 10 out of 29 publications