Carboxyfluorescein Dye Uptake to Measure Connexin-mediated Hemichannel Activity in Cultured Cells.

Potter, Joe, Price, Gareth, Cliff, Chelsy , Williams, Bethany, Hills, Claire and Squires, Paul (2021) Carboxyfluorescein Dye Uptake to Measure Connexin-mediated Hemichannel Activity in Cultured Cells. Manual. Bio-Protocol, Bio-Protocol.

Full content URL:

Full text not available from this repository.

Item Type:Paper or Report (Manual)
Item Status:Live Archive


Connexins are membrane bound proteins that facilitate direct and local paracrine mediated cell-to-cell communication through their ability to oligomerise into hexameric hemichannels. When neighbouring channels align, they form gap-junctions that provide a direct route for information transfer between cells. In contrast to intact gap junctions, which typically open under physiological conditions, undocked hemichannels have a low open probability and mainly open in response to injury. Hemichannels permit the release of small molecules and ions (approximately 1kDa) into the local intercellular environment, and excessive expression/activity has been linked to a number of disease conditions. Carboxyfluorescein dye uptake measures functional expression of hemichannels, where increased hemichannel activity/function reflects increased loading. The technique relies on the uptake of a membrane-impermeable fluorescent tracer through open hemichannels, and can be used to compare channel activity between cell monolayers cultured under different conditions, e.g. control versus disease. Other techniques, such as biotinylation and electrophysiology can measure cell surface expression and hemichannel open probability respectively, however, carboxyfluorescein uptake provides a simple, rapid and cost-effective method to determine hemichannel activity in vitro in multiple cell types.

Keywords:Connexins, Hemichannels, Carboxyfluorescein, Dye uptake
Subjects:C Biological Sciences > C130 Cell Biology
B Subjects allied to Medicine > B120 Physiology
Divisions:College of Science > School of Life Sciences
Related URLs:
ID Code:43924
Deposited On:15 Feb 2021 10:45

Repository Staff Only: item control page