Abstract
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.
MeSH Terms
Animals
Calcium/pharmacology
Calcium Channels/drug effects,physiology
Cesium/pharmacology
Heart/physiology
Kinetics
Lipid Bilayers
Membrane Potentials
Models, Biological
Muscle Proteins/drug effects,physiology
Muscle, Skeletal/physiology
Patch-Clamp Techniques
Phosphatidylcholines
Phosphatidylethanolamines
Phosphatidylserines
Rabbits
Ryanodine Receptor Calcium Release Channel
Sarcoplasmic Reticulum/drug effects,physiology
Sheep
Solutions
Surface Properties
Chemicals
Calcium Channels
Lipid Bilayers
Muscle Proteins
Phosphatidylcholines
Phosphatidylethanolamines
Phosphatidylserines
Ryanodine Receptor Calcium Release Channel
Solutions
1-palmitoyl-2-oleoylphosphatidylethanolamine
Cesium
1-palmitoyl-2-oleoylglycero-3-phosphoserine
Calcium
1-palmitoyl-2-oleoylphosphatidylcholine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Laver D R
Division of Neuroscience, John Curtin School of Medical Research, Australian National University, Canberra, Australia. derek.laver@anu.edu.au
Curtis B A
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