Measuring extracellular ion gradients from single channels with ion-selective microelectrodes.
Under many different conditions activated plasma membrane ion channels give rise to changes in the extracellular concentration of the permeant ion(s). The magnitude and duration of these changes are dependent on the electrochemical driving force(s) on the permeant ion(s) as well as conductance, open time, and channel density. We have modeled the change in the extracellular [K+] due to efflux through Ca2+-activated K+ channels, mSlo, to determine the range of parameters that would give rise to measurable signals in the surrounding media. Subsequently we have used extracellular, K+-selective microelectrodes to monitor localized changes in [K+]ext due to efflux through mSlo channels expressed in Xenopus oocytes. The rapid changes in [K+] show a close fit with the predicted model when the time response of the ion-selective microelectrode is taken into account, providing proof of the concept. Measurement of the change in extracellular ion concentration with ion-selective microelectrodes provides a noninvasive means for functional mapping of channel location and density, as well as characterizing the properties of ion channels in the plasma membrane.
Messerli
Mark A
MA
Corson
Erica D
ED
Smith
Peter J S
PJ
D000818:Animals; D002478:Cells, Cultured; D045604:Extracellular Fluid / Q000378:metabolism*; D015640:Ion Channel Gating / Q000502:physiology*; D007477:Ions; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits / Q000502:physiology*; D008839:Microelectrodes; D009865:Oocytes / Q000502:physiology*; D018408:Patch-Clamp Techniques / Q000295:instrumentation* / Q000379:methods; D012680:Sensitivity and Specificity; D014982:Xenopus laevis
D007477:Ions; C465449:Kcnma1 protein, mouse; D051037:Large-Conductance Calcium-Activated Potassium Channel alpha Subunits