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@article{1380301,
author = {Upham, BL and Sovadinová, Iva and Babica, Pavel},
article_location = {CAMBRIDGE},
article_number = {118},
doi = {http://dx.doi.org/10.3791/54281},
keywords = {Cellular Biology; Issue 118; gap junctional intercellular communication; scalpel load-fluorescent dye transfer; in vitro toxicology; biomarker; cell signaling; signal transduction; cancer; chemoprevention},
language = {eng},
issn = {1940-087X},
journal = {JOVE-JOURNAL OF VISUALIZED EXPERIMENTS},
title = {Gap Junctional Intercellular Communication: A Functional Biomarker to Assess Adverse Effects of Toxicants and Toxins, and Health Benefits of Natural Products},
volume = {2016},
year = {2016}
}
TY - JOUR
ID - 1380301
AU - Upham, BL - Sovadinová, Iva - Babica, Pavel
PY - 2016
TI - Gap Junctional Intercellular Communication: A Functional Biomarker to Assess Adverse Effects of Toxicants and Toxins, and Health Benefits of Natural Products
JF - JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
VL - 2016
IS - 118
SP - "e54281"-"e54289"
EP - "e54281"-"e54289"
PB - JOURNAL OF VISUALIZED EXPERIMENTS
SN - 1940087X
KW - Cellular Biology
KW - Issue 118
KW - gap junctional intercellular communication
KW - scalpel load-fluorescent dye transfer
KW - in vitro toxicology
KW - biomarker
KW - cell signaling
KW - signal transduction
KW - cancer
KW - chemoprevention
N2 - 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.
ER -
UPHAM, BL, Iva SOVADINOVÁ a Pavel BABICA. Gap Junctional Intercellular Communication: A Functional Biomarker to Assess Adverse Effects of Toxicants and Toxins, and Health Benefits of Natural Products. \textit{JOVE-JOURNAL OF VISUALIZED EXPERIMENTS}. CAMBRIDGE: JOURNAL OF VISUALIZED EXPERIMENTS, roč.~2016, č.~118, s.~''e54281''-''e54289'', 9 s. ISSN~1940-087X. doi:10.3791/54281. 2016.
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