Insulin promoter-driven Gaussia luciferase-based insulin secretion biosensor assay for discovery of β-cell glucose-sensing pathways.
High 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.
Department of Phamacology, UT Southwestern Medical Center, Dallas, TX 75235, USA.
Michael A
MA
Kalwat
Department of Phamacology, UT Southwestern Medical Center, Dallas, TX 75235, USA.
Chonlarat
C
Wichaidit
Department of Phamacology, UT Southwestern Medical Center, Dallas, TX 75235, USA.
Alejandra Y
AY
Nava Garcia
Department of Biochemistry, UT Southwestern Medical Center, Dallas, TX 75235, USA.
Melissa K
MK
McCoy
Department of Phamacology, UT Southwestern Medical Center, Dallas, TX 75235, USA.
Kathleen
K
McGlynn
Department of Biochemistry, UT Southwestern Medical Center, Dallas, TX 75235, USA.
In Hyun
IH
Hwang
Department of Biochemistry, UT Southwestern Medical Center, Dallas, TX 75235, USA.
John B
JB
MacMillan
Department of Biochemistry, UT Southwestern Medical Center, Dallas, TX 75235, USA.
Bruce A
BA
Posner
Department of Phamacology, UT Southwestern Medical Center, Dallas, TX 75235, USA.
Melanie H
MH
Cobb
Gaussia; high-throughput screening; insulin; pancreatic islet beta cell marine natural products.