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Related Experiment Videos

ATP enhances catecholamine uptake into PC12 cells

A Eshleman1, C Dunigan, A Shamoo

  • 1Department of Pharmacology and Experimental Therapeutics, University of Maryland School of Medicine, Baltimore 21201, USA.

Life Sciences
|January 1, 1995
PubMed
Summary

Adenosine triphosphate (ATP) significantly boosts catecholamine uptake in PC12 cells, acting as an allosteric modulator. This ATP-enhanced uptake may lead to novel cocaine antagonists.

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Pharmacology

Background:

  • Catecholamine transporters are crucial for neurotransmission.
  • Adenosine triphosphate (ATP) is an energy currency with potential signaling roles.
  • PC12 cells are a widely used model for neuronal function.

Purpose of the Study:

  • To investigate the effect of ATP on catecholamine uptake in PC12 cells.
  • To elucidate the mechanism and conditions under which ATP influences uptake.
  • To explore the potential therapeutic implications of ATP's effect.

Main Methods:

  • Catecholamine uptake assays in PC12 cells using dopamine and norepinephrine.
  • Experiments conducted in bicarbonate-buffered Krebs Ringer solution.
  • Varying concentrations of ATP and other nucleotides were tested.

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  • Uptake sensitivity to temperature, Na(+), and inhibitors was assessed.

    • Main Results:

    • ATP significantly enhanced catecholamine uptake in a dose-dependent manner (maximal at 1-3 mM).
    • ATP increased Vmax by 3-5 fold for dopamine and norepinephrine, without altering Km.
    • ATP-stimulated uptake was temperature- and Na(+)-dependent and buffer-specific (bicarbonate vs. HEPES).
    • GTP and UTP showed similar enhancement effects; ATP metabolites did not.
    • ATP-modulated uptake exhibited altered sensitivity to inhibitors.

    Conclusions:

    • ATP acts as an allosteric modulator of the catecholamine transporter.
    • The findings suggest a novel regulatory mechanism for catecholamine transport.
    • A stable ATP analog could potentially serve as a cocaine antagonist.