Efficacy of the gap junction blocker carbenoxolone was visualized by transfer of Lucifer Yellow as previously described (Opsahl and Rivedal, 2000 (link)). Briefly, epithelial monolayers were wounded in the presence of 1.0% Lucifer Yellow (Thermo Fisher Scientific) in PBS. Tissues were incubated for 5 min at 37°C prior to visualization. Gap junctions were blocked using carbenoxolone disodium (50 µM; Sigma) applied 24 h prior to performing the assay. Plots of the fluorescence intensity as a function of distance from the scratch were created in ImageJ.
Carbenoxolone disodium
Manufactured by Merck Group
Sourced in Germany, United States
Carbenoxolone disodium is a chemical compound used in laboratory settings. It is a synthetic derivative of glycyrrhizic acid, and its primary function is as a corrosion inhibitor in various laboratory applications.
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3 protocols using carbenoxolone disodium
1
Visualizing Gap Junction Blockade
2
Investigating Small Molecule Efficacy
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Tetraethylthiuram disulfide (DIS), Temozolomide (TMZ) and Carbenoxolone disodium (CBX) were obtained from Sigma-Aldrich (Steinheim, Germany) and dissolved in DMSO (DIS and TMZ) or water (CBX). For in vivo experiments CBX was replaced by INI-0602 (Wako Pure Chemical Ind., Ltd., Japan).
3
Effects of Carbenoxolone on SCN Cell Activity
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In six experiments, we tested the effects of i.c.v. administration of the gap junction blocker carbenoxolone (carbenoxolone disodium; Sigma, St Louis, MO, USA) on the activity of an SCN cell. carbenoxolone (2 μl of 10 or 100 mg ml−1 solution in artificial cerebrospinal fluid) was administered into the third ventricle over 2–3 min via a cannula inserted into the ventricle through the median eminence, 3 mm caudal to the recording site. These experiments were all conducted in rats maintained on a normal light cycle.
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