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Related Experiment Videos

Protein kinase C as mediator of arachidonic acid-induced decrease of neuronal M current

H Schmitt1, H Meves

  • 1I. Physiologisches Institut, Universität des Saarlandes, Homburg-Saar, Germany.

Pflugers Archiv : European Journal of Physiology
|October 1, 1993
PubMed
Summary

Arachidonic acid reduces the M current (IM) in NG108-15 cells. This effect is largely mediated by protein kinase C (PKC) activation, as shown by inhibition experiments.

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Biochemistry

Background:

  • The M current (IM) is a non-inactivating potassium current crucial for neuronal excitability.
  • NG108-15 cells, a hybridoma cell line, express the M current.
  • Arachidonic acid is known to modulate ion channel function.

Purpose of the Study:

  • To investigate the mechanism by which arachidonic acid affects the M current in NG108-15 cells.
  • To determine if protein kinase C (PKC) activation mediates the effect of arachidonic acid on IM.
  • To characterize the impact of PKC inhibition on arachidonic acid-induced changes in IM.

Main Methods:

  • Electrophysiological recordings (voltage-clamp) to measure the M current (IM).
  • Application of arachidonic acid (5-25 microM) to NG108-15 cells.

Related Experiment Videos

  • Use of a specific protein kinase C (PKC) inhibitor peptide (PKC 19-31) to block PKC activity.
  • Pharmacological activation of PKC using phorbol 12,13-dibutyrate (PDB).

    • Main Results:

    • Arachidonic acid (5-25 microM) decreased the M current (IM) in NG108-15 cells, sometimes with a transient increase.
    • A PKC inhibitor peptide (1 microM) completely prevented the inhibitory effect of phorbol 12,13-dibutyrate (PDB) on IM, confirming effective PKC inhibition.
    • In the presence of the PKC inhibitor peptide, the reduction of IM by arachidonic acid (25 microM) was significantly attenuated.
    • PKC inhibition also shifted the voltage dependence of the M conductance (gM(V)) to more negative potentials.

    Conclusions:

    • Arachidonic acid modulates the M current (IM) in NG108-15 cells.
    • The reduction in IM by arachidonic acid is significantly mediated by the activation of protein kinase C (PKC).
    • PKC plays a key role in regulating the M current in these neuroblastoma x glioma hybrid cells.