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Updated: May 17, 2026
09:47
An Iodide-Yellow Fluorescent Protein-Gap Junction-Intercellular Communication Assay
Published on: February 1, 2019
A high-throughput assay for connexin 43 (Cx43, GJA1) gap junctions using codon-optimized aequorin
Nazia Haq1, David Grose, Emma Ward
1Biological Reagents and Assay Development, GlaxoSmithKline R&D, Stevenage, United Kingdom.
Assay and Drug Development Technologies
|October 11, 2012
Summary
This study presents a new high-throughput assay to screen for drugs targeting gap junctions (GJs) by measuring calcium transfer between cells. This method facilitates the discovery of novel therapeutic modulators for GJ-related conditions.
Area of Science:
- Cellular Biology
- Pharmacology
- Biophysics
Background:
- Gap junctions (GJs) facilitate intercellular communication via connexin proteins.
- Modulators of GJs show therapeutic potential but require careful study.
- Existing assays for GJ function are limited in throughput.
Purpose of the Study:
- To develop a high-throughput assay for screening GJ modulators.
- To characterize the functionality of connexin 43 (Cx43) mediated calcium transfer.
- To enable drug discovery for conditions involving GJ dysfunction.
Main Methods:
- Exogenous expression of human Cx43 in HeLa cells.
- A 384-well assay measuring luminescence from aequorin-based calcium indicators.
- Stimulation of calcium signaling via adrenergic receptors or TRPV1 channels.
Main Results:
- Demonstrated dose-dependent calcium transfer between Cx43-expressing cells.
- Confirmed Cx43 dependency and inhibition by GJ blockers.
- Identified calcium as a likely permeant ion through GJs.
Conclusions:
- The developed assay is suitable for high-throughput screening of GJ modulators.
- This method can identify compounds affecting Cx43 channel activity.
- The assay advances drug discovery for GJ-related diseases.
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