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Insulin Promoter-Driven Gaussia Luciferase-Based Insulin Secretion Biosensor Assay for Discovery of β‑Cell Glucose-Sensing Pathways
journal contribution
posted on 2016-10-12, 00:00 authored by Michael
A. Kalwat, Chonlarat Wichaidit, Alejandra Y. Nava Garcia, Melissa K. McCoy, Kathleen McGlynn, In Hyun Hwang, John B. MacMillan, Bruce
A. Posner, Melanie H. CobbHigh
throughput screening of insulin secretion is intractable with
current methods. We developed a secreted insulin–luciferase
system (Ins-GLuc) in β cells that is rapid, inexpensive, and
amenable to 96- and 384-well formats. We treated stable Ins-GLuc-expressing
MIN6 cells overnight with 6298 marine natural product fractions. The
cells were then washed to remove media and chemicals, followed by
stimulation with glucose in the diazoxide paradigm. These conditions
allowed the discovery of many insulin secretion suppressors and potentiators.
The mechanisms of action of these natural products must be long-lasting
given the continuance of secretory phenotypes in the absence of chemical
treatment. We anticipate that these natural products and their target
pathways will lead to a greater understanding of glucose-stimulated
insulin secretion.
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chemical treatmenttarget pathwaysβ cellsdiazoxide paradigmproduct fractions6298 marineinsulin secretion suppressorssecretory phenotypesglucose-stimulated insulin secretionInsulin Promoter-Driven Gaussia Luciferase-Based Insulin Secretion Biosensor AssayIns-GLuc-expressing MIN 6 cellsinsulin secretion
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