UPHAM, BL, Iva SOVADINOVÁ and Pavel BABICA. Gap Junctional Intercellular Communication: A Functional Biomarker to Assess Adverse Effects of Toxicants and Toxins, and Health Benefits of Natural Products. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS. CAMBRIDGE: JOURNAL OF VISUALIZED EXPERIMENTS, 2016, vol. 2016, No 118, p. "e54281"-"e54289", 9 pp. ISSN 1940-087X. Available from: https://dx.doi.org/10.3791/54281.
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Basic information
Original name Gap Junctional Intercellular Communication: A Functional Biomarker to Assess Adverse Effects of Toxicants and Toxins, and Health Benefits of Natural Products
Authors UPHAM, BL (840 United States of America), Iva SOVADINOVÁ (203 Czech Republic, belonging to the institution) and Pavel BABICA (203 Czech Republic, belonging to the institution).
Edition JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, CAMBRIDGE, JOURNAL OF VISUALIZED EXPERIMENTS, 2016, 1940-087X.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 30304 Public and environmental health
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 1.232
RIV identification code RIV/00216224:14310/16:00094239
Organization unit Faculty of Science
Doi http://dx.doi.org/10.3791/54281
UT WoS 000397846600008
Keywords in English Cellular Biology; Issue 118; gap junctional intercellular communication; scalpel load-fluorescent dye transfer; in vitro toxicology; biomarker; cell signaling; signal transduction; cancer; chemoprevention
Tags AKR, rivok
Changed by Changed by: Ing. Andrea Mikešková, učo 137293. Changed: 11/5/2017 15:16.
Abstract
This protocol describes a scalpel loading-fluorescent dye transfer (SL-DT) technique that measures intercellular communication through gap junction channels, which is a major intercellular process by which tissue homeostasis is maintained. Interruption of gap junctional intercellular communication (GJIC) by toxicants, toxins, drugs, etc. has been linked to numerous adverse health effects. Many genetic-based human diseases have been linked to mutations in gap junction genes. The SL-DT technique is a simple functional assay for the simultaneous assessment of GJIC in a large population of cells. The assay involves pre-loading cells with a fluorescent dye by briefly perturbing the cell membrane with a scalpel blade through a population of cells. The fluorescent dye is then allowed to traverse through gap junction channels to neighboring cells for a designated time. The assay is then terminated by the addition of formalin to the cells. The spread of the fluorescent dye through a population of cells is assessed with an epifluorescence microscope and the images are analyzed with any number of morphometric software packages that are available, including free software packages found on the public domain. This assay has also been adapted for in vivo studies using tissue slices from various organs from treated animals. Overall, the SL-DT assay can serve a broad range of in vitro pharmacological and toxicological needs, and can be potentially adapted for high throughput set-up systems with automated fluorescence microscopy imaging and analysis to elucidate more samples in a shorter time.
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