Measurement of Ensemble TRPV1 Ion Channel Currents Using Droplet Bilayers
نویسندگان
چکیده
Electrophysiological characterization of ion channels is useful for elucidation of channel function as well as quantitative assessment of pharmaceutical effects on ion channel conductance. We used droplet bilayers to measure ensemble ion channel currents from membrane preparations made from TRPV1-expressing HEK cells. Conductance measurements showed rectification, activation by acid and capsaicin, and inhibition by capsazepine, SB 452533, and JNJ 17293212. We also quantitatively measured concentration-dependent inhibition of channel conductance through determination of capsazepine IC50 in agreement with previously published studies using patch clamp. These results, combined with the reduced apparatus and material requirements of droplet bilayers, indicate that this platform could be used for study of other physiologically relevant ion channels.
منابع مشابه
Activation of the mechanosensitive ion channel MscL by mechanical stimulation of supported Droplet-Hydrogel bilayers
The droplet on hydrogel bilayer (DHB) is a novel platform for investigating the function of ion channels. Advantages of this setup include tight control of all bilayer components, which is compelling for the investigation of mechanosensitive (MS) ion channels, since they are highly sensitive to their lipid environment. However, the activation of MS ion channels in planar supported lipid bilayer...
متن کاملBp 7.2 Mon 17:45 P3 Scanning Fluorescence Correlation Spectroscopy on Mem- Branes — Curvature-coupled Protein Diffusion in a Fluctuating Model Membrane — Elucidating Structure and Domain Formation of Biomimetic Influence of Tension on Coarse-grained Model Membranes —
Biological lipid membranes are predominantly asymmetric. In the plasma membranes of eukaryotic cells, for example, there is an abundance of phosphatidylcholine and sphingomyelin in the outer leaflet while aminophospholipids are primarily in the cytosolic leaflet. The biological importance of asymmetric lipid bilayers has motivated many studies using model systems, such as planar bilayers, suppo...
متن کاملA Portable Lipid Bilayer System for Environmental Sensing with a Transmembrane Protein
This paper describes a portable measurement system for current signals of an ion channel that is composed of a planar lipid bilayer. A stable and reproducible lipid bilayer is formed in outdoor environments by using a droplet contact method with a micropipette. Using this system, we demonstrated that the single-channel recording of a transmembrane protein (alpha-hemolysin) was achieved in the f...
متن کاملLactate is a potent inhibitor of the capsaicin receptor TRPV1
Tissue ischemia results in an accumulation of lactate and local or systemic lactic acidosis. In nociceptive sensory neurons, lactate was reported to sensitize or activate the transient receptor potential ion channel TRPA1 and acid-sensing ion channels (ASICs). However, it is unclear how lactate modulates the TRPV1 regarded as the main sensor for acidosis in sensory neurons. In this study we inv...
متن کاملCharging the Quantum Capacitance of Graphene with a Single Biological Ion Channel
The interaction of cell and organelle membranes (lipid bilayers) with nanoelectronics can enable new technologies to sense and measure electrophysiology in qualitatively new ways. To date, a variety of sensing devices have been demonstrated to measure membrane currents through macroscopic numbers of ion channels. However, nanoelectronic based sensing of single ion channel currents has been a ch...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
دوره 10 شماره
صفحات -
تاریخ انتشار 2015