J 2016

Gap Junctional Intercellular Communication: A Functional Biomarker to Assess Adverse Effects of Toxicants and Toxins, and Health Benefits of Natural Products

UPHAM, BL, Iva SOVADINOVÁ and Pavel BABICA

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

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

30304 Public and environmental health

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 1.232

RIV identification code

RIV/00216224:14310/16:00094239

Organization unit

Faculty of Science

DOI

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

Změněno: 11/5/2017 15:16, Ing. Andrea Mikešková

Abstract

V originále

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.