Single-channel properties of a G-protein-coupled inward rectifier potassium channel in brain neurons.
1. In cultured rat locus coeruleus neurons, somatostatin or met-enkephalin induces an inwardly rectifying K+ conductance. This inward rectifier was analyzed at the single-channel level. 2. Using the inside-out patch-clamp, guanosine 5'-triphosphate (GTP) application to the cytoplasmic side in the presence of somatostatin or met-enkephalin in the pipette produced a large increase in channel activity, which disappeared on switching from GTP to guanosine 5'-diphosphate. 3. The unitary conductance was approximately 30 pS at -95 mV with an extracellular K+ concentration of 156 mM and an intracellular K+ concentration of 124 mM at 23 degrees C. The channel showed burst behavior, and the closed time histogram was fit by two exponentials, with the fast time constant being 0.4 ms. The burst time histogram was also fit by two exponentials, with time constants of 0.24 and 2.0 ms (at 10 nM somatostatin). When the somatostatin concentration was changed from 500 to 1 nM, the kinetic behavior of the channel did not change, except that the open probability of the patch was decreased. 4. The current-voltage relation of the unitary channel current showed inward rectification. The reversal potential coincided with the K+ equilibrium potential, and it shifted according to a change in the K+ equilibrium potential. 5. In the presence of external somatostatin, the application of guanosine 5'-O-(3-thiotriphosphate) to the cytoplasmic side induced an irreversible activation of this channel. 6. These results indicate that this K+ channel is the microscopic counterpart of the somatostatin- or met-enkephalin-induced inwardly rectifying K+ current in whole cell recording, and that the channel is activated by a G protein without a diffusible second messenger. Thus this channel is identified as a neuronal G-protein-coupled inward rectifier K+ channel. 7. Analysis of the burst behavior, based on a close-close-open kinetic model, revealed that there are at least four states in the K+ channel, a short gap, a longer closing, a short opening, and a long opening, and that the neuronal inward rectifier is activated at faster rates than the atrial inward rectifier.
Grigg
J J
JJ
Department of Anatomy and Cell Biology, University of Illinois at Chicago 60612, USA.
Kozasa
T
T
Nakajima
Y
Y
Nakajima
S
S
D000328:Adult; D000818:Animals; D001921:Brain / Q000502:physiology*; D046508:Culture Techniques; D004744:Enkephalin, Methionine / Q000502:physiology; D019204:GTP-Binding Proteins / Q000502:physiology*; D006160:Guanosine Triphosphate / Q000502:physiology; D006801:Humans; D008125:Locus Coeruleus / Q000502:physiology; D008564:Membrane Potentials; D009474:Neurons / Q000502:physiology; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000502:physiology*; D051381:Rats; D013004:Somatostatin / Q000502:physiology; D009435:Synaptic Transmission / Q000502:physiology*
D013004:Somatostatin; D004744:Enkephalin, Methionine; D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; D011188:Potassium