Carbachol-induced phosphoinositide turnover in NCB-20 cells
NCB-20 cells (fetal Chinese hamster brain cell x neuroblastoma hybrids) have been shown to contain a variety of neurotransmitter receptors. The authors now report that this cloned cell line also contains acetylcholne receptors which are linked to phospholipase C. Confluent cell cultures were preincubated with /sup 3/H-myo-inositol to label endogenous phosphoinositide (PI) and the accumulation of a PI metabolite, inositol monophosphate (IP/sub 1/), was measured in the presence of LiCl. Carbachol increased IP/sub 1/), accumulation be more than 400% with a EC/sub 50/ of about 50 ..mu..M. Acetylcholine and muscarine were also effective, whereas oxotremorine and McN-A-343 were weak in both potency and efficacy. The carbachol-induced IP/sub 1/ accumulation was completely blocked by atropine (Ki approx. 0.6 nM) and pirenzepine (Ki approx. 15 nM). The presence of KCl was not required for the carbachol-induced effect. The formation of inositol bis- and triphosphate was also increased carbachol; these increases occurred earlier but were of much smaller magnitude. Pretreatment of cells with 4 ..beta..-phorbol dibutyrate or 4 ..beta..-phorbol myristate acetate was found to attenuate the carbachol-induced formation of IP/sub 1/ (IC/sub 50/ in the low nanomolar concentration ranges), however 4 ..beta..-phorbol, the biologically inactive phorbol ester, was ineffective in causing this attenuation. These results suggest a feedback inhibition of PI turnover in NCB-20 cells through the action of protein kinase C.
- Research Organization:
- National Institute of Mental Health, Washington, DC
- OSTI ID:
- 5275603
- Report Number(s):
- CONF-8604222-; TRN: 86-030561
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
ACETYLCHOLINE
RECEPTORS
INOSITOL
BIOLOGICAL ACCUMULATION
PARASYMPATHOMIMETICS
BIOLOGICAL EFFECTS
PHORBOL ESTERS
BIOCHEMICAL REACTION KINETICS
BRAIN
CELL CULTURES
CLONE CELLS
FETUSES
HAMSTERS
LIPASE
METABOLITES
NERVES
PHOSPHATES
PHOSPHOTRANSFERASES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
AMINES
AMMONIUM COMPOUNDS
ANIMALS
AUTONOMIC NERVOUS SYSTEM AGENTS
BODY
CARBOHYDRATES
CARBOXYLESTERASES
CARCINOGENS
CENTRAL NERVOUS SYSTEM
DRUGS
ENZYMES
ESTERASES
ESTERS
HYDROLASES
INOSITOLS
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MAMMALS
MEMBRANE PROTEINS
MONOSACCHARIDES
NERVOUS SYSTEM
NEUROREGULATORS
ORGANIC COMPOUNDS
ORGANS
OXYGEN COMPOUNDS
PHOSPHORUS COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PROTEINS
QUATERNARY COMPOUNDS
REACTION KINETICS
RODENTS
SACCHARIDES
TRANSFERASES
VERTEBRATES
560301* - Chemicals Metabolism & Toxicology- Cells- (-1987)
550201 - Biochemistry- Tracer Techniques