Acetylcholine secretion in the human cell strain LA-N-2
The authors have studied the synthesis and release of acetylcholine (ACh) in human neuroblastoma cells, LA-N-2. In cells cultured for 4 days in nutrient medium containing 7-700 ..mu..M choline, the cell content as well as the amounts of ACh spontaneously released into the medium increased with increasing substrate concentration. Half-maximal intracellular ACh levels were reached at 40 /sup +/M medium choline. Incubation of LA-N-2 cells for 1 hr with (/sup 3/H)choline and subsequent purification of the radioactive species by HPLC, showed incorporation of radioactive choline into ACh in a saturable manner with an apparent Km of 82 +/- 17 ..mu..M and a Vmax of 1.4 +/- 0.1 nmol/mg protein/hr. ACh secretion by LA-N-2 cells is stimulated by 1. elevated concentrations of extracellular K/sup +/ but not Cs/sup +/; 2. a sodium channel agonist, veratridine, an effect blocked by tetrodotoxin and 3. A cholinergic agonist, carbachol, an effect blocked by atropine. LA-N-2 cells thus have retained important features of differentiated neuronal cells and offer a model system for studies on the molecular mechanisms of cholinergic function.
- Research Organization:
- Massachusetts Institute of Technology, Cambridge
- OSTI ID:
- 6179829
- Report Number(s):
- CONF-870644-; TRN: 87-034016
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 46:6; Conference: 78. annual meeting of the American Society of Biological Chemists conference, Philadelphia, PA, USA, 7 Jun 1987
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ACETYLCHOLINE
BIOSYNTHESIS
SECRETION
CHOLINE
FRACTIONATION
BIOLOGICAL MODELS
CELL CULTURES
LIQUID COLUMN CHROMATOGRAPHY
MAN
NERVE CELLS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
TUMOR CELLS
ALCOHOLS
AMINES
AMMONIUM COMPOUNDS
ANIMAL CELLS
ANIMALS
AUTONOMIC NERVOUS SYSTEM AGENTS
CHROMATOGRAPHY
DRUGS
ESTERS
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIPOTROPIC FACTORS
MAMMALS
NEUROREGULATORS
ORGANIC COMPOUNDS
PARASYMPATHOMIMETICS
PRIMATES
QUATERNARY COMPOUNDS
SEPARATION PROCESSES
SOMATIC CELLS
SYNTHESIS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques