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An Iodide-Yellow Fluorescent Protein-Gap Junction-Intercellular Communication Assay
Published on: February 1, 2019
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An Iodide-Yellow Fluorescent Protein-Gap Junction-Intercellular Communication Assay.
1College of Pharmacy, Yonsei Institute of Pharmaceutical Sciences, Yonsei University.
Journal of Visualized Experiments : Jove
|February 19, 2019
Summary
A new assay uses iodide and fluorescent proteins to measure gap junction communication. This method enables high-throughput screening for drug discovery and toxicology studies.
Area of Science:
- Cell Biology
- Biochemistry
Background:
- Gap junctions (GJs) facilitate intercellular communication by allowing molecule diffusion between adjacent cells.
- Physiological and pathological roles of GJs necessitate high-throughput screening (HTS) assays for identifying GJ modulators.
- Existing methods may not be suitable for rapid drug discovery and toxicology assessments.
Purpose of the Study:
- To develop and validate a novel, high-throughput assay for measuring gap junction intercellular communication (GJIC).
- To provide a reliable method for identifying modulators of GJ activity relevant to drug discovery and toxicology.
Main Methods:
- A novel iodide-yellow fluorescent protein-gap junction-intercellular communication (I-YFP-GJIC) assay was developed.
- The assay utilizes engineered donor and acceptor cells expressing an iodide transporter (SLC26A4) and a YFP variant, respectively.
- GJIC activity is quantified by measuring YFP fluorescence quenching upon iodide influx and GJ-mediated diffusion.
Main Results:
- The I-YFP-GJIC assay demonstrated reliable and rapid measurement of GJ activity.
- The YFP fluorescence quenching rate directly correlates with the extent of GJIC.
- The assay is suitable for high-throughput screening applications.
Conclusions:
- The I-YFP-GJIC assay is a robust tool for HTS of GJ modulators.
- This assay facilitates drug discovery and toxicological evaluations by assessing GJ function.
- The described protocol using LN215 cells provides a standardized method for GJIC assessment.
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