Developing High-Throughput Assays to Analyze and Screen Electrophysiological Phenotypes

Methods Mol Biol. 2018:1787:235-252. doi: 10.1007/978-1-4939-7847-2_18.

Abstract

Ion channels represent nearly a quarter of all targets that currently available medications modulate, and their dysfunction underlies increasing number of human diseases. Functional analysis of ion channels have traditionally been a bottleneck in large-scale analyses. Recent technological breakthroughs in automated planar electrophysiology have democratized the technique to enable high-throughput patch clamping at scale. In this chapter, we describe the methodology to perform a phenotypic screen on voltage-gated calcium channels across many different genetic coding variations and against small-molecule modulators. We first describe the procedures to establish inducible heterologous ion channel expression in HEK293 cells, where each cell incorporates one copy of a target protein cDNA-a step that is critical for producing stable and consistent expression of ion channels. We then describe the experimental and analytical methods for analyzing the function of ion channels using high-throughput planar electrophysiology.

Keywords: Electrophysiology; High throughput; Ion channels; Patch clamp; Phenotypic screen; Planar electrophysiology; Voltage-gated calcium channels.

MeSH terms

  • Calcium Channels / genetics
  • Calcium Channels / metabolism
  • Data Interpretation, Statistical
  • Drug Discovery
  • Electrophysiological Phenomena*
  • Gene Expression
  • HEK293 Cells
  • High-Throughput Screening Assays*
  • Humans
  • Ion Channel Gating
  • Patch-Clamp Techniques
  • Workflow

Substances

  • Calcium Channels