The alpha9alpha10 nicotinic acetylcholine receptor is permeable to and is modulated by divalent cations.
The native cholinergic receptor that mediates synaptic transmission between olivocochlear fibers and outer hair cells of the cochlea is permeable to Ca(2+) and is thought to be composed of both the alpha 9 and the alpha 10 cholinergic nicotinic subunits. The aim of the present work was to study the permeability of the recombinant alpha 9 alpha 10 nicotinic acetylcholine receptor to Ca(2+), Ba(2+) and Mg(2+) and its modulation by these divalent cations. Experiments were performed, by the two-electrode voltage-clamp technique, in Xenopus laevis oocytes injected with alpha 9 and alpha 10 cRNA. The relative divalent to monovalent cation permeability was high ( approximately 10) for Ca(2+), Ba(2+) and Mg(2+). Currents evoked by acetylcholine (ACh) were potentiated by either Ca(2+) or Ba(2+) up to 500 microM but were blocked by higher concentrations of these cations. Potentiation by Ca(2+) was voltage-independent, whereas blockage was stronger at hyperpolarized than at depolarized potentials. Mg(2+) did not potentiate but it blocked ACh-evoked currents (IC(50)=0.38 mM). In the absence of Ca(2+), the EC(50) for ACh was higher (48 microM) than that obtained with 1.8 mM Ca(2+) (14.3 microM), suggesting that potentiation by Ca(2+) involves changes in the apparent affinity of the alpha 9 alpha 10 receptor for ACh.
Instituto de Investigaciones en Ingeniería Genética y Biología Molecular (CONICET-UBA), Vuelta de Obligado 2490, 1428 Buenos Aires, Argentina.
Noelia
N
Weisstaub
Douglas E
DE
Vetter
Ana Belén
AB
Elgoyhen
Eleonora
E
Katz
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002413:Cations, Divalent / Q000494:pharmacology; D004594:Electrophysiology; D005260:Female; D018072:Hair Cells, Auditory, Outer / Q000378:metabolism; D066298:In Vitro Techniques; D008274:Magnesium / Q000494:pharmacology; D009865:Oocytes / Q000378:metabolism; D010539:Permeability; D021122:Protein Subunits; D051381:Rats; D011978:Receptors, Nicotinic / Q000737:chemistry / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D009435:Synaptic Transmission; D014982:Xenopus laevis
D002413:Cations, Divalent; C498541:Chrna9 protein, rat; D021122:Protein Subunits; D011978:Receptors, Nicotinic; D011994:Recombinant Proteins; D008274:Magnesium; D002118:Calcium