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Importance of voltage-dependent inactivation in N-type calcium channel regulation by G-proteins

  • Norbert Weiss
  • Abir Tadmouri
  • Mohamad Mikati
  • Michel Ronjat
  • Michel De WaardEmail author
Ion Channels

Abstract

Direct regulation of N-type calcium channels by G-proteins is essential to control neuronal excitability and neurotransmitter release. Binding of the \({\text{G}}_{{\beta \gamma }} \) dimer directly onto the channel is characterized by a marked current inhibition (“ON” effect), whereas the pore opening- and time-dependent dissociation of this complex from the channel produce a characteristic set of biophysical modifications (“OFF” effects). Although G-protein dissociation is linked to channel opening, the contribution of channel inactivation to G-protein regulation has been poorly studied. Here, the role of channel inactivation was assessed by examining time-dependent G-protein de-inhibition of Cav2.2 channels in the presence of various inactivation-altering β subunit constructs. G-protein activation was produced via μ-opioid receptor activation using the DAMGO agonist. Whereas the “ON” effect of G-protein regulation is independent of the type of β subunit, the “OFF” effects were critically affected by channel inactivation. Channel inactivation acts as a synergistic factor to channel activation for the speed of G-protein dissociation. However, fast inactivating channels also reduce the temporal window of opportunity for G-protein dissociation, resulting in a reduced extent of current recovery, whereas slow inactivating channels undergo a far more complete recovery from inhibition. Taken together, these results provide novel insights on the role of channel inactivation in N-type channel regulation by G-proteins and contribute to the understanding of the physiological consequence of channel inactivation in the modulation of synaptic activity by G-protein coupled receptors.

Keywords

N-type calcium channel Cav2.2 subunit G-protein G-protein coupled receptor μ-opioid receptor inactivation β subunit 

Abbreviations

GI

G-protein inhibition

GPCR

G-protein coupled receptor

DAMGO

(d-Ala2,Me-Phe4,glycinol5)-enkephalin

rMOR

rat μ-opioid receptor

PCR

polymerase chain reaction

RI

recovery from inhibition

NS

non-statistically significant

Notes

Acknowledgements

We thank Dr. Pierre Charnet and Dr. Yasuo Mori for providing the cDNAs encoding the rat μ-opioid receptor and the rabbit Cav2.2 channel, respectively. We are indebted to Dr. Anne Feltz, Dr. Lubica Lacinova, Dr. Michel Vivaudou, and Dr. Eric Hosy for critical evaluation of this work. We thank Sandrine Geib for her contribution to the CD8–β1b construct.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Norbert Weiss
    • 1
    • 3
    • 4
  • Abir Tadmouri
    • 1
    • 3
    • 4
  • Mohamad Mikati
    • 2
  • Michel Ronjat
    • 1
    • 3
    • 4
  • Michel De Waard
    • 1
    • 3
    • 4
    Email author
  1. 1.Laboratoire Canaux Calciques, Fonctions et PathologiesInserm U607, CEAGrenoble Cedex 09France
  2. 2.Department of PediatricsAmerican University of Beirut Medical CenterBeirutLebanon
  3. 3.Commissariat à l’Energie AtomiqueGrenobleFrance
  4. 4.Université Joseph FourierGrenobleFrance

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