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Related Experiment Videos

Extracellular ATP stimulates volume decrease in Necturus red blood cells.

D B Light1, T L Capes, R T Gronau

  • 1Department of Biology, Ripon College, Ripon, Wisconsin 54971, USA. LightD@Ripon.edu

The American Journal of Physiology
|September 14, 1999
PubMed
Summary
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Extracellular ATP promotes regulatory volume decrease in mudpuppy red blood cells by stimulating calcium-dependent potassium efflux. This finding reveals a novel role for external ATP in cell volume regulation.

Area of Science:

  • Cell Physiology
  • Ion Transport
  • Red Blood Cell Biology

Background:

  • Regulatory Volume Decrease (RVD) is crucial for cell survival under osmotic stress.
  • The role of extracellular adenosine triphosphate (ATP) in mediating RVD in non-mammalian red blood cells remains largely unexplored.
  • Understanding ATP's function in red blood cells (RBCs) can provide insights into cellular homeostasis and signaling pathways.

Purpose of the Study:

  • To investigate whether extracellular ATP stimulates regulatory volume decrease (RVD) in Necturus maculosus red blood cells.
  • To elucidate the specific ion transport mechanisms and signaling pathways involved in ATP-mediated RVD.

Main Methods:

  • Hemolytic index measurements to assess osmotic fragility.
  • Electronic cell volume measurements using a Coulter counter during hypotonic shock.

Related Experiment Videos

  • Whole-cell patch-clamp electrophysiology to record ion currents.
  • Pharmacological manipulation using ATP, ATP scavengers (hexokinase, apyrase), channel blockers (glibenclamide, gadolinium), and ionophores (A-23187, gramicidin).

    • Main Results:

    • Extracellular ATP (50 microM) decreased osmotic fragility and enhanced cell volume recovery during hypotonic shock.
    • ATP-scavenging enzymes and K+ channel blockers (glibenclamide, gadolinium) inhibited RVD, effects reversed by ATP.
    • Patch-clamp studies revealed ATP-stimulated whole-cell currents, dependent on Ca2+ and linked to K+ efflux, with evidence of ATP efflux during swelling.

    Conclusions:

    • Extracellular ATP acts as a potent stimulator of regulatory volume decrease in Necturus maculosus red blood cells.
    • ATP-mediated RVD involves a Ca2+-dependent pathway leading to K+ efflux through specific ion channels.
    • Cell swelling may trigger ATP efflux, suggesting a feedback mechanism in cell volume regulation.