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Protein kinase C and regulatory volume decrease in mudpuppy red blood cells

D B Light1, M R Adler, J K Ter Beest

  • 1Department of Biology, Ripon College, Ripon, WI 54971, USA.

The Journal of Membrane Biology
|December 5, 1998
PubMed
Summary
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Protein kinase C (PKC) regulates potassium (K+) efflux during cell volume regulation in mudpuppy red blood cells. Inhibiting PKC reduces K+ loss and cell volume recovery, indicating its role in this process.

Area of Science:

  • Cellular Physiology
  • Biochemistry
  • Membrane Transport

Background:

  • Regulatory Volume Decrease (RVD) is a critical cellular process for maintaining cell volume homeostasis.
  • Potassium (K+) efflux is a key mechanism driving RVD in many cell types, including red blood cells.
  • The role of protein kinase C (PKC) in regulating ion transport during RVD remains incompletely understood.

Purpose of the Study:

  • To investigate the involvement of protein kinase C (PKC) in stimulating K+ efflux during RVD in Necturus maculosus red blood cells (RBCs).
  • To identify the specific conductive pathways regulated by PKC during the RVD process.

Main Methods:

  • Utilized osmotic fragility tests and cell volume measurements (Coulter counter, hematocrit) to assess RVD.
  • Employed specific inhibitors of protein kinases, including general inhibitors (H-7) and PKC antagonists (bisindolylmaleimide I/II, chelerythrine).

Related Experiment Videos

  • Applied patch clamp techniques to measure whole-cell conductance changes during cell swelling.

    • Main Results:

    • Inhibition of PKC with specific antagonists (bis I, bis II, chelerythrine) and a general kinase inhibitor (H-7) increased osmotic fragility and decreased RVD.
    • The effects of PKC inhibition on RVD and osmotic fragility were reversed by gramicidin, confirming a link to K+ efflux.
    • Patch clamp data revealed that H-7 and PKC inhibitors reduced a K+-conductive pathway activated during cell swelling.

    Conclusions:

    • Protein kinase C (PKC) plays a significant role in regulating K+ efflux during regulatory volume decrease (RVD) in Necturus maculosus red blood cells.
    • A conductive pathway mediating K+ loss during RVD is, at least partially, regulated by PKC.
    • These findings elucidate a novel regulatory mechanism for ion transport involved in cell volume control.