Kinetics and autoradiography of high affinity uptake of serotonin by primary astrocyte cultures

Katz, D M; Kimelberg, H K

Title: Kinetics and autoradiography of high affinity uptake of serotonin by primary astrocyte cultures

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Abstract

Primary astrocyte cultures prepared from the cerebral cortices of neonatal rats showed significant accumulation of serotonin (5-hydroxytryptamine; (/sup 3/H)-5-HT). At concentrations in the range of 0.01 to 0.7 microM (/sup 3/H)-5-HT, this uptake was 50 to 85% Na+ dependent and gave a Km of 0.40 +/- 0.11 microM (/sup 3/H)-5-HT and a Vmax of 6.42 +/- 0.85 (+/- SEM) pmol of (/sup 3/H)-5-HT/mg of protein/4 min for the Na+-dependent component. In the absence of Na+ the uptake was nonsaturable. Omission of the monoamine oxidase inhibitor pargyline markedly reduced the Na+-dependent component of (/sup 3/H)-5-HT uptake but had a negligible effect on the Na+-independent component. This suggest significant oxidative deamination of serotonin after it has been taken up by the high affinity system, followed by release of its metabolite. The authors estimated that this system enabled the cells to concentrate (/sup 3/H)-5-HT up to 44-fold at an external (/sup 3/H)-5-HT concentration of 10(-7) M. Inhibition of (/sup 3/H)-5-HT uptake by a number of clinically effective antidepressants was also consistent with a specific high affinity uptake mechanism for 5-HT, the order of effectiveness of inhibition being chlorimipramine greater than fluoxetine greater than imipramine = amitriptyline greater than desmethylimipramine greater than iprindole greatermore »« less

Authors:: Katz, D M; Kimelberg, H K

Publication Date:: Mon Jul 01 00:00:00 EDT 1985

Research Org.:: Albany Medical College, NY

OSTI Identifier:: 5098408

Resource Type:: Journal Article

Journal Name:: J. Neurosci.; (United States)

Additional Journal Information:: Journal Volume: 7

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; ANTIDEPRESSANTS; BIOLOGICAL EFFECTS; SEROTONIN; BIOCHEMICAL REACTION KINETICS; UPTAKE; AFFINITY; ANIMAL CELLS; AUTORADIOGRAPHY; CATIONS; CELL CULTURES; CEREBRAL CORTEX; METABOLITES; NEONATES; OXIDOREDUCTASES; RATS; SODIUM; TRACER TECHNIQUES; TRITIUM COMPOUNDS; ALKALI METALS; AMINES; ANIMALS; AROMATICS; AUTONOMIC NERVOUS SYSTEM AGENTS; AZAARENES; AZOLES; BODY; BRAIN; CENTRAL NERVOUS SYSTEM; CENTRAL NERVOUS SYSTEM AGENTS; CEREBRUM; CHARGED PARTICLES; DRUGS; ELEMENTS; ENZYMES; HETEROCYCLIC COMPOUNDS; HYDROXY COMPOUNDS; INDOLES; IONS; ISOTOPE APPLICATIONS; KINETICS; LABELLED COMPOUNDS; MAMMALS; METALS; NERVOUS SYSTEM; NEUROREGULATORS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; ORGANS; PSYCHOTROPIC DRUGS; PYRROLES; RADIOPROTECTIVE SUBSTANCES; REACTION KINETICS; RODENTS; SYMPATHOMIMETICS; TRYPTAMINES; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques

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                    Katz, D M, and Kimelberg, H K. Kinetics and autoradiography of high affinity uptake of serotonin by primary astrocyte cultures.  United States: N. p., 1985. 
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                    Katz, D M, & Kimelberg, H K. Kinetics and autoradiography of high affinity uptake of serotonin by primary astrocyte cultures.  United States.

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                    Katz, D M, and Kimelberg, H K. 1985.  
        "Kinetics and autoradiography of high affinity uptake of serotonin by primary astrocyte cultures".  United States.

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@article{osti_5098408,

  title        = {Kinetics and autoradiography of high affinity uptake of serotonin by primary astrocyte cultures},

  author       = {Katz, D M and Kimelberg, H K},

  abstractNote = {Primary astrocyte cultures prepared from the cerebral cortices of neonatal rats showed significant accumulation of serotonin (5-hydroxytryptamine; (/sup 3/H)-5-HT). At concentrations in the range of 0.01 to 0.7 microM (/sup 3/H)-5-HT, this uptake was 50 to 85% Na+ dependent and gave a Km of 0.40 +/- 0.11 microM (/sup 3/H)-5-HT and a Vmax of 6.42 +/- 0.85 (+/- SEM) pmol of (/sup 3/H)-5-HT/mg of protein/4 min for the Na+-dependent component. In the absence of Na+ the uptake was nonsaturable. Omission of the monoamine oxidase inhibitor pargyline markedly reduced the Na+-dependent component of (/sup 3/H)-5-HT uptake but had a negligible effect on the Na+-independent component. This suggest significant oxidative deamination of serotonin after it has been taken up by the high affinity system, followed by release of its metabolite. The authors estimated that this system enabled the cells to concentrate (/sup 3/H)-5-HT up to 44-fold at an external (/sup 3/H)-5-HT concentration of 10(-7) M. Inhibition of (/sup 3/H)-5-HT uptake by a number of clinically effective antidepressants was also consistent with a specific high affinity uptake mechanism for 5-HT, the order of effectiveness of inhibition being chlorimipramine greater than fluoxetine greater than imipramine = amitriptyline greater than desmethylimipramine greater than iprindole greater than mianserin. Uptake of (/sup 3/H)-5-HT was dependent on the presence of Cl- as well as Na+ in the medium, and the effect of omission of both ions was nonadditive. Varying the concentration of K+ in the media from 1 to 50 mM had a limited effect on (/sup 3/H)-5-HT uptake.},

  doi          = {},

  url          = {https://www.osti.gov/biblio/5098408},
  journal      = {J. Neurosci.; (United States)},
number       = ,

  volume       = 7,

  place        = {United States},

  year         = {Mon Jul 01 00:00:00 EDT 1985},

  month        = {Mon Jul 01 00:00:00 EDT 1985}

}

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