Functional coupling between human E-type Ca2+ channels and mu opioid receptors expressed in Xenopus oocytes.
Neuronal alpha1E Ca2+ channels were expressed in Xenopus laevis oocytes alone and in combination with the mu opioid receptor. Macroscopic currents were recorded under voltage clamp conditions. The stimulation of the morphine receptor by the synthetic [D-Ala2,N-Me-Phe4,Gly-ol5] enkephalin (DAMGO) produced a 20% reduction in the alpha1E ionic current. This effect was associated with a large change in the decay phase of the Ba2+ current. The effect of 1 microM DAMGO was fully antagonized by the universal mu opioid receptor antagonist naloxone and by the selective antagonist beta-funaltrexamine. The ionic current inhibition induced by DAMGO was partially recovered by preceding strong depolarizations. The injection of the catalytic subunit of pertussis toxin (A-protomer) abolished the effect of DAMGO, suggesting the involvement of a GTP binding protein in the alpha1E modulation. The coexpression of the regulatory beta2a Ca2a channel subunit, together with the alpha1E subunit and the mu opioid receptor, prevented the reduction of the ionic current following the receptor stimulation with DAMGO, whereas the coexpression with the beta3 subunit reduced by approximately 50% the modulatory effect of DAMGO. The effect produced by the stimulation of the opioid receptor could be mimicked by coexpressing the alpha1E channel with the G-protein betagamma subunits.
Ottolia
M
M
Department of Anesthesiology, University of California Los Angeles, 90095-1778, USA.
Platano
D
D
Qin
N
N
Noceti
F
F
Birnbaumer
M
M
Toro
L
L
Birnbaumer
L
L
Stefani
E
E
Olcese
R
R
D000701:Analgesics, Opioid / Q000494:pharmacology*; D000818:Animals; D015220:Calcium Channels / Q000187:drug effects* / Q000378:metabolism; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D004745:Enkephalins / Q000494:pharmacology*; D019204:GTP-Binding Proteins / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects; D009294:Narcotics / Q000494:pharmacology; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D037342:Pertussis Toxin; D017450:Receptors, Opioid, mu / Q000187:drug effects* / Q000378:metabolism; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D014981:Xenopus / Q000235:genetics / Q000378:metabolism
D000701:Analgesics, Opioid; D015220:Calcium Channels; D004745:Enkephalins; D009294:Narcotics; D017450:Receptors, Opioid, mu; D010566:Virulence Factors, Bordetella; D020875:Enkephalin, Ala(2)-MePhe(4)-Gly(5)-; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins