Novel KCNQ2 channel activators discovered using fluorescence-based and automated patch-clamp-based high-throughput screening techniques.

ABSTRACT:

Aim

To establish an improved, high-throughput screening techniques for identifying novel KCNQ2 channel activators.

Methods

KCNQ2 channels were stably expressed in CHO cells (KCNQ2 cells). Thallium flux assay was used for primary screening, and 384-well automated patch-clamp IonWorks Barracuda was used for hit validation. Two validated activators were characterized using a conventional patch-clamp recording technique.

Results

From a collection of 80 000 compounds, the primary screening revealed a total of 565 compounds that potentiated the fluorescence signals in thallium flux assay by more than 150%. When the 565 hits were examined in IonWorks Barracuda, 38 compounds significantly enhanced the outward currents recorded in KCNQ2 cells, and were confirmed as KCNQ2 activ

SUBMITTER: Yue JF 

PROVIDER: S-EPMC4722984 | biostudies-literature | 2016 Jan

REPOSITORIES: biostudies-literature