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

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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.

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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

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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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Authors and Affiliations

  1. Laboratoire Canaux Calciques, Fonctions et Pathologies, Inserm U607, CEA, 17 rue des Martyrs, 38054, Grenoble Cedex 09, France

    Norbert Weiss, Abir Tadmouri, Michel Ronjat & Michel De Waard

  2. Department of Pediatrics, American University of Beirut Medical Center, Beirut, Lebanon

    Mohamad Mikati

  3. Commissariat à l’Energie Atomique, Grenoble, France

    Norbert Weiss, Abir Tadmouri, Michel Ronjat & Michel De Waard

  4. Université Joseph Fourier, Grenoble, France

    Norbert Weiss, Abir Tadmouri, Michel Ronjat & Michel De Waard

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  1. Norbert Weiss
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  2. Abir Tadmouri
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  3. Mohamad Mikati
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  4. Michel Ronjat
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  5. Michel De Waard
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Correspondence to Michel De Waard.

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Weiss, N., Tadmouri, A., Mikati, M. et al. Importance of voltage-dependent inactivation in N-type calcium channel regulation by G-proteins. Pflugers Arch - Eur J Physiol 454, 115–129 (2007). https://doi.org/10.1007/s00424-006-0184-0

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