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Kinetic analysis of the interaction between noradrenaline and Na+ in neuronal uptake: Kinetic evidence for CO-transport

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Summary

  1. 1.

    Vasa deferentia obtained from reserpine-pretreated rats were incubated (under conditions of inhibition of both monoamine oxidase and catechol O-methyltransferase) in medium containing various concentrations of 3H-(−)noradrenaline (1.25–30.25 μmol·l−1) and Na+ (0–143 mmol·l−1; isosmolality maintained by Tris+). Initial rates of neuronal uptake (v i ) were determined in each single vas from the difference between the uptake of noradrenaline occurring in the absence and that occurring in the presence of 100 μmol·l−1 cocaine.

  2. 2.

    The uptake of noradrenaline observed after exposure to cocaine was virtually identical with that observed after incubation in Na+-free medium (containing or not containing cocaine). Under these experimental conditions, 70% of the uptake was due to extracellular distribution of the amine, and not only this part of uptake, but also the remaineder was linearly related to the noradrenaline concentration in the medium.

  3. 3.

    The neuronal uptake of noradrenaline showed saturation with increasing concentrations of noradrenaline or Na+. When determined at several fixed concentrations of Na+ (or noradrenaline), the plots of 1/v i vs. 1/[noradrenaline] (or 1/[Na+]) were all linear and intersected at a common point to the left of the ordinate and above the abscissa. Increases in the fixed concentration of Na+ (or noradrenaline) progressively increased the apparent V max and progressively decreased the apparent K m of the system for noradrenaline (or Na+). Moreover, the vertical intercept (1/apparent V max) and the slope (apparent ratio of K m /V max) of the Lineweaver-Burk plots were linearly related to the reciprocal of the concentration of the “fixed” substrate.

  4. 4.

    Thus, the neuronal uptake mechanism exhibits the kinetic properties of a two-substrate sequential reaction in which both noradrenaline and Na+ (1:1) must bind to the carrier for transport of noradrenaline to occur and in which noradrenaline and Na+ act as mutually cooperative co-substrates.

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Authors and Affiliations

  1. Institut für Pharmakologie und Toxikologie der Universität Würzburg, Versbacher Strasse 9, D-8700, Würzburg, Federal Republic of Germany

    S. Sammet & K.-H. Graefe

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  1. S. Sammet
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  2. K.-H. Graefe
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This study was supported by the Deutsche Forschungsgemeinschaft (Gr. 490/4)

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Sammet, S., Graefe, KH. Kinetic analysis of the interaction between noradrenaline and Na+ in neuronal uptake: Kinetic evidence for CO-transport. Naunyn-Schmiedeberg's Arch. Pharmacol. 309, 99–107 (1979). https://doi.org/10.1007/BF00501216

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  • DOI: https://doi.org/10.1007/BF00501216

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