You are here

Methods Mol Biol DOI:10.1007/978-1-4939-7847-2_18

Developing High-Throughput Assays to Analyze and Screen Electrophysiological Phenotypes.

Publication TypeJournal Article
Year of Publication2018
AuthorsPan, JQ, Baez-Nieto, D, Allen, A, Wang, H-R, Cottrell, JR
JournalMethods Mol Biol
Volume1787
Pages235-252
Date Published2018
ISSN1940-6029
KeywordsCalcium Channels, Data Interpretation, Statistical, Drug Discovery, Electrophysiological Phenomena, Gene Expression, HEK293 Cells, High-Throughput Screening Assays, Humans, Ion Channel Gating, Patch-Clamp Techniques, Workflow
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.

DOI10.1007/978-1-4939-7847-2_18
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/29736723?dopt=Abstract

Alternate JournalMethods Mol. Biol.
PubMed ID29736723
Grant ListR01 MH115045 / MH / NIMH NIH HHS / United States