Surface potentials measure ion concentrations near lipid bilayers during rapid solution changes.
We describe a puffing method for changing solutions near one surface of lipid bilayers that allows simultaneous measurement of channel activity and extent of solution change at the bilayer surface. Ion adsorption to the lipid headgroups and screening of the bilayer surface charge by mobile ions provided a convenient probe for the ionic composition of the solution at the bilayer surface. Rapid ionic changes induced a shift in bilayer surface potential that generated a capacitive transient current under voltage-clamp conditions. This depended on the ion species and bilayer composition and was accurately described by the Stern-Gouy-Chapman theory. The time course of solute concentrations during solution changes could also be modeled by an exponential exchange of bath and puffing solutions with time constants ranging from 20 to 110 ms depending on the flow pressure. During changes in [Cs+] and [Ca2+] (applied separately or together) both the mixing model and capacitive currents predicted [Cs+] and [Ca2+] transients consistent with those determined experimentally from: 1) the known Cs(+)-dependent conductance of open ryanodine receptor channels and 2) the Ca(2+)-dependent gating of ryanodine receptor Ca2+ channels from cardiac and skeletal muscle.
Laver
D R
DR
Division of Neuroscience, John Curtin School of Medical Research, Australian National University, Canberra, Australia. derek.laver@anu.edu.au
Curtis
B A
BA
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D015220:Calcium Channels / Q000187:drug effects / Q000502:physiology*; D002586:Cesium / Q000494:pharmacology; D006321:Heart / Q000502:physiology*; D007700:Kinetics; D008051:Lipid Bilayers; D008564:Membrane Potentials; D008954:Models, Biological; D009124:Muscle Proteins / Q000187:drug effects / Q000502:physiology*; D018482:Muscle, Skeletal / Q000502:physiology*; D018408:Patch-Clamp Techniques; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D011817:Rabbits; D019837:Ryanodine Receptor Calcium Release Channel; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000502:physiology*; D012756:Sheep; D012996:Solutions; D013499:Surface Properties
D015220:Calcium Channels; D008051:Lipid Bilayers; D009124:Muscle Proteins; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D019837:Ryanodine Receptor Calcium Release Channel; D012996:Solutions; C057561:1-palmitoyl-2-oleoylphosphatidylethanolamine; D002586:Cesium; C050294:1-palmitoyl-2-oleoylglycero-3-phosphoserine; D002118:Calcium; C028694:1-palmitoyl-2-oleoylphosphatidylcholine