Insulin promoter-driven Gaussia luciferase-based insulin secretion biosensor assay for discovery of ?-cell glucose-sensing pathways.

ABSTRACT: High throughput screening of insulin secretion is intractable with current methods. We developed a secreted insulin-luciferase system (Ins-GLuc) in ? 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.

SUBMITTER: Kalwat MA 

PROVIDER: S-EPMC5089812 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature