Activation of a common potassium channel in molluscan neurones by glutamate, dopamine and muscarinic agonist.
1. The potassium currents evoked in isolated and identified neurones of molluscan pedal ganglia by either glutamate, dopamine or the muscarinic agonist F-2268 were investigated using voltage and patch clamp techniques. 2. Potassium currents induced by either dopamine or F-2268 could be blocked by pertussis toxin, as well as by a prolonged intracellular injection of the G protein inhibitor, GDP-beta-S. Loading the neurones with the G protein activator, GppNHp, on the other hand, induced a potassium current. This current was not additive to the currents evoked by agonist application. 3. Intracellular injection of the calcium buffer BAPTA failed to affect any of the agonist-induced currents, although it effectively blocked the after-hyperpolarization following directly evoked action potentials. 4. The activity of the potassium channels seen in cell-attached patches was greatly enhanced by application to the bath of either glutamate, dopamine, or F-2268. 5. The only effect of an addition of agonists to the bath was to increase the open probability (Po) of the K+ channel already active in the control conditions. The identity of the spontaneously active and agonist-activated channels was concluded from the identity of their channel conductances, rectification properties and current amplitudes. 6. Phorbol-12,13-dibutyrate, when applied to the bath, induced an increase in open time and caused an increase in Po, as did the agonists. Staurosporine completely prevented changes of Po induced by the phorbol ester but not those induced by the agonists. 7. The same inwardly rectifying potassium channel may be opened by both the receptor-linked G protein (with glutamate, dopamine, F-2268) and by protein kinase C (with phorbol ester) activation. 8. Strong evidence was obtained against the involvement of any known secondary messenger systems (formation of nucleotides, phosphoinositide turnover and subsequent activation of protein kinase C, formation of nitric oxide, metabolism of arachidonic acid) in the transduction mechanism of F-2268-, dopamine- and glutamate-induced responses. 9. Since none of the known secondary messenger systems seems to affect the activation by agonists applied to receptors outside the patch of channels located under the patch electrode, it appears that some as yet undescribed linking system must exist that could connect the spatially separated receptor-G protein complex and the potassium channel.
Bolshakov VYu
I. M. Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St Petersburg.
Gapon
S A
SA
Katchman
A N
AN
Magazanik
L G
LG
D000818:Animals; D004148:Dioxolanes / Q000494:pharmacology; D004298:Dopamine / Q000494:pharmacology; D005071:Evoked Potentials / Q000187:drug effects; D019204:GTP-Binding Proteins / Q000378:metabolism; D005971:Glutamates / Q000494:pharmacology; D018698:Glutamic Acid; D007700:Kinetics; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D010277:Parasympathomimetics / Q000494:pharmacology; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism; D011954:Receptors, Dopamine / Q000187:drug effects / Q000378:metabolism; D017981:Receptors, Neurotransmitter / Q000187:drug effects / Q000378:metabolism; D015290:Second Messenger Systems; D015398:Signal Transduction; D012908:Snails / Q000378:metabolism*
D004148:Dioxolanes; D005971:Glutamates; D010277:Parasympathomimetics; D015221:Potassium Channels; D011954:Receptors, Dopamine; D017981:Receptors, Neurotransmitter; D015240:Phorbol 12,13-Dibutyrate; D018698:Glutamic Acid; C028858:methamilane methiodide; D011493:Protein Kinase C; D019204:GTP-Binding Proteins; D004298:Dopamine