Localization-dependent activity of the Kv2.1 delayed-rectifier K+ channel.
The Kv2.1 K(+) channel is highly expressed throughout the brain, where it regulates excitability during periods of high-frequency stimulation. Kv2.1 is unique among Kv channels in that it targets to large surface clusters on the neuronal soma and proximal dendrites. These clusters also form in transfected HEK cells. Following excessive excitatory stimulation, Kv2.1 declusters with an accompanying 20- to 30-mV hyperpolarizing shift in the activation threshold. Although most Kv2.1 channels are clustered, there is a pool of Kv2.1 resident outside of these domains. Using the cell-attached patch clamp technique, we investigated the hypothesis that Kv2.1 activity varies as a function of cell surface location. We found that clustered Kv2.1 channels do not efficiently conduct K(+), whereas the nonclustered channels are responsible for the high threshold delayed rectifier K(+) current typical of Kv2.1. Comparison of gating and ionic currents indicates only 2% of the surface channels conduct, suggesting that the clustered channels still respond to membrane potential changes. Declustering induced via either actin depolymerization or alkaline phosphatase treatment did not increase whole-cell currents. Dephosphorylation resulted in a 25-mV hyperpolarizing shift, whereas actin depolymerization did not alter the activation midpoint. Taken together, these data demonstrate that clusters do not contain high threshold Kv2.1 channels whose voltage sensitivity shifts upon declustering; nor are they a reservoir of nonconducting channels that are activated upon release. On the basis of these findings, we propose unique roles for the clustered Kv2.1 that are independent of K(+) conductance.
O'Connell
Kristen M S
KM
Department of Biomedical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
Loftus
Robert
R
Tamkun
Michael M
MM
D000465:Algorithms; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015640:Ion Channel Gating / Q000502:physiology*; D008564:Membrane Potentials; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000378:metabolism*; D045663:Quantum Dots; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D051669:Shab Potassium Channels / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014162:Transfection
D011993:Recombinant Fusion Proteins; D051669:Shab Potassium Channels; D049452:Green Fluorescent Proteins; D011188:Potassium