Parallel Recordings of Transmembrane hERG Channel Currents Based on Solvent-Free Lipid Bilayer Microarray.

Ryusuke Miyata,Daisuke Tadaki,Ayumi Hirano-Iwata,Maki Komiya,Shun Araki,Madoka Sato,Teng Ma,Daichi Yamaura,Michio Niwano,Hideaki Yamamoto

doi:10.3390/mi12010098

Ryusuke Miyata, Daisuke Tadaki + Show 8 more

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https://doi.org/10.3390/mi12010098

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Abstract

The reconstitution of ion-channel proteins in artificially formed bilayer lipid membranes (BLMs) forms a well-defined system for the functional analysis of ion channels and screening of the effects of drugs that act on these proteins. To improve the efficiency of the BLM reconstitution system, we report on a microarray of stable solvent-free BLMs formed in microfabricated silicon (Si) chips, where micro-apertures with well-defined nano- and micro-tapered edges were fabricated. Sixteen micro-wells were manufactured in a chamber made of Teflon®, and the Si chips were individually embedded in the respective wells as a recording site. Typically, 11 to 16 BLMs were simultaneously formed with an average BLM number of 13.1, which corresponded to a formation probability of 82%. Parallel recordings of ion-channel activities from multiple BLMs were successfully demonstrated using the human ether-a-go-go-related gene (hERG) potassium channel, of which the relation to arrhythmic side effects following drug treatment is well recognized.

Highlights

The cell membrane is composed of a bilayer lipid membrane (BLM), a self-assembled structure of phospholipid membrane molecules, and membrane proteins embedded in the bilayer lipid membranes (BLMs) [1]
We presented a 16-site BLM microarray system based on microfabricated
We succeeded in the simultaneous formation of solvent-free BLMs over 16 micro-apertures fabricated in Si chips

Summary

Introduction

The cell membrane is composed of a bilayer lipid membrane (BLM), a self-assembled structure of phospholipid membrane molecules, and membrane proteins embedded in the BLM [1]. Ion-channel proteins function as gated pores that permit ion fluxes across membranes that have a considerably high resistance (>1 GΩ). Electrical recording of transmembrane ion-channel currents is one of the most efficient methods for characterizing the channel functions and screening the effects of drugs acting on ion-channel proteins [6,7]. Fluorometric measurement of transmembrane ion fluxes using liposomes [8] or BLMs suspended on micro- and nano-cavity arrays [9,10] is useful for high-throughput recordings of channel functions. Only electrical recording can control and clamp the transmembrane voltage, which is necessary for the functional analysis of voltage-gated ion channels. The patch-clamping method has been the gold standard for the analysis of drug actions on ion Micromachines 2021, 12, 98.

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Discussion

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