Regulation of muscarinic acetylcholine receptors in the 1321N1 human astrocytoma cell line
The binding of muscarinic agonists, partial agonists and antagonists to muscarinic receptors of 1321N1 human astrocytoma cells was studied. Binding was studied in both intact cells and cell lysates. Partial agonists and antagonists exhibited similar apparent affinities in intact cell competition binding assays with either the lipophilic radioligand ({sup 3}H)QNB or the hydrophilic radioligand ({sup 3}H)NMS. In contrast, full agonists exhibited markedly lower apparent affinities in intact cells with ({sup 3}H)QNB than with ({sup 3}H)NMS. Treatment of cells with antimycin A to deplete intracellular ATP prevented agonist-induced internalization of muscarinic receptors as assessed by sucrose density gradient assays of receptor subcellular distribution. In ATP-depleted cells, the apparent affinities of full agonists vs ({sup 3}H)QNB were markedly higher. The apparent affinities of partial agonists and of antagonists were unaffected by ATP depletion. In other studies, the effects of the protein kinase C activator phorbol 12-myristate, 13-acetate (PMA) on muscarinic receptor downregulation and internalization in 1321N1 cells were determined. PMA alone did not induce muscarinic receptor downregulation but instead decreased both the rate and final extent of downregulation induced by the agonist carbachol. The specificity of other protein kinase C activators for inhibiting carbachol-induced downregulation indicated involvement of protein kinase C. Furthermore, the protein kinase C inhibitor staurosporine prevented the inhibitory effect of PMA on downregulation. However, staurosporine did not inhibit agonist-induced downregulation.
- Research Organization:
- Missouri Univ., Columbia, MO (United States)
- OSTI ID:
- 5545274
- Resource Relation:
- Other Information: Thesis (Ph. D.)
- Country of Publication:
- United States
- Language:
- English
Similar Records
Ligand binding and functional characterization of muscarinic acetylcholine receptors on the TE671/RD human cell line
Characterization of the muscarinic cholinoceptors in the human detrusor
Related Subjects
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ACETYLCHOLINE
RECEPTORS
PHORBOL ESTERS
BIOLOGICAL EFFECTS
AFFINITY
BIOCHEMICAL REACTION KINETICS
INHIBITION
LIGANDS
MAN
PARASYMPATHOLYTICS
PHOSPHOTRANSFERASES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
TUMOR CELLS
AMINES
AMMONIUM COMPOUNDS
ANIMAL CELLS
ANIMALS
AUTONOMIC NERVOUS SYSTEM AGENTS
CARCINOGENS
DRUGS
ENZYMES
ESTERS
HYDROGEN COMPOUNDS
ISOTOPE APPLICATIONS
KINETICS
MAMMALS
MEMBRANE PROTEINS
NEUROREGULATORS
ORGANIC COMPOUNDS
PARASYMPATHOMIMETICS
PHOSPHORUS-GROUP TRANSFERASES
PRIMATES
PROTEINS
QUATERNARY COMPOUNDS
REACTION KINETICS
TRANSFERASES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques
560300 - Chemicals Metabolism & Toxicology