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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 347 (1993), S. 371-378 
    ISSN: 1432-1912
    Keywords: Human amnion FL cells ; Noradrenaline ; Uptake2 ; O-methylation ; Papaverine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The uptake and metabolism of 3H-noradrenaline has been examined in the FL cell-line derived originally from human amnion. Cell cultures metabolised 3H-noradrenaline (1.0 μmol/l) to 3H-normetanephrine and, to a lesser extent, to metabolites (not distinguished) of the O-methylated deaminated fraction; primary deaminated metabolites were not detected. 3(H-normetanephrine formation a) was not saturable in the noradrenaline concentration range 0.2–150 μmol/l, b was decreased to 20%–30% of control levels by uptake2 inhibitors (O-methylisoprenaline, 20 and 100 μmol/l; cimetidine, 10 μmol/l; hydrocortisone, 200 μmol/l) and c, was almost insensitive to uptake1 inhibitors (cocaine, 30 μmol/l; desipramine, 3 μmol/l). Uptake of noradrenaline was manifested after 30 minutes as a 6-fold increase in the cell content of the amine following inhibition of catechol-O-methyl transferase, either alone or in conjunction with inhibition of monoamine oxidase. Uptake was decreased maximally to 40% of control levels by O-methylisoprenaline. IC50 values for inhibition of the O-methylisoprenaline-sensitive component of uptake were (in μmol/l): corticosterone (0.3), papaverine (1.1), O-methylisoprenaline (3.0), cimetidine (6.0), (−)noradrenaline (460), and tetraethylammonium (2230). Except for the last agent, for which a comparative value is not available, the IC50's are in good agreement with those for inhibition of uptake2 in the Caki-1 cell-line reported by other investigators. The component of uptake resistant to O-methylisoprenaline was depressed by papaverine (a 50% decrease at 50 μmol/l), but was not affected by the other uptake2 inhibitors or by cocaine (30 μmol/l). It is concluded that the FL cell possesses an extraneuronal metabolising system very similar to the system in tissues such as heart and smooth muscle where transport of noradrenaline into the cell by uptake2 is followed by rapid O-methylation via catechol-O-methyl transferase. The only difference appears to be the absence of saturation of 3H-normetanephrine formation in the FL cell at low micromolar concentrations of 3H-noradrenaline. The presence of a second uptake process is suggested by the inhibitory effect of papaverine on uptake resistant to O-methylisoprenaline; lack of effect of cocaine implies that this second process is not uptake,.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Naunyn-Schmiedeberg's archives of pharmacology 346 (1992), S. 166-172 
    ISSN: 1432-1912
    Keywords: Dental pulp ; Noradrenaline ; Extraneuronal uptake1 ; O-methylation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The extraneuronal removal and disposition of noradrenaline in rabbit dental pulp was examined in view of earlier evidence that the tissue possessed an extra-neuronal uptake process resembling neuronal uptake1. Pulp, which had been depleted of sympathetic nerves by homolateral superior cervical ganglionectomy, was incubated in vitro with 3H-noradrenaline in low concentrations (0.025 or 0.18 μmol/l). When the metabolising enzymes (monoamine oxidase, catechol-O-methyl transferase) were active, 3H retention by the denervated pulp, as indicated by the 3H content after the tissue had been washed for 30 min following incubation with 3H-noradrenaline, was less than 30% of that of the innervated pulp. When the enzymes were inhibited, retention rose to approximately 30% of that of the innervated pulp. Analysis of the time course of the 3H efflux indicated that the 3H-noradrenaline in the denervated pulp had accumulated in a single compartment characterised by a t1/2 for efflux of several hours. Accumulation did not occur under Na+-free conditions, and was inhibited by desipramine (IC50 〈 0.03 μmol/l) and by substrates of neuronal uptake1. Mean IC50, values of the latter were very similar to those for inhibition of neuronal uptake1 and comprised (in μmol/l): (+)amphetamine (0.29), dopamine (0.31), tyramine (0.39), (−)noradrenaline (0.70), (−)adrenaline (1.50), 5-hydroxytryptamine (20) and bretylium (35). Uptake2 inhibitors were less active (O-methyl isoprenaline, IC50 = 60 μmol/l) than uptake1 inhibitors, or were without inhibitory effects at the concentrations tested (hydrocortisone, 210 μmol/l; 2-methoxy oestrone, 10 μmol/l). The effects of Na+ omission, of (+)amphetamine, and of O-methylisoprenaline on 3H-normetanephrine formation (measured in the absence of catechol-O-methyl transferase inhibition) matched their effects on 3H-noradrenaline accumulation. The results provide strong support for the presence in rabbit dental pulp of extraneuronal uptake1 which is linked with catechol-O-methyl transferase in the removal of noradrenaline.
    Type of Medium: Electronic Resource
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