title: Novel systems for dynamically assessing insulin action in live cells reveals heterogeneity in the insulin response.
abstract: Regulated GLUT4 trafficking is a key action of insulin. Quantitative stepwise analysis of this process provides a powerful tool for pinpointing regulatory nodes that contribute to insulin regulation and insulin resistance. We describe a novel GLUT4 construct and workflow for the streamlined dissection of GLUT4 trafficking; from simple high throughput screens to high resolution analyses of individual vesicles. We reveal single cell heterogeneity in insulin action highlighting the utility of this approach - each cell displayed a unique and highly reproducible insulin response, implying that each cell is hard-wired to produce a specific output in response to a given stimulus. These data highlight that the response of a cell population to insulin is underpinned by extensive heterogeneity at the single cell level. This heterogeneity is pre-programmed within each cell and is not the result of intracellular stochastic events.
authors.affiliation: Diabetes and Obesity Research Program, The Garvan Institute of Medical Research, Sydney, NSW, 2010, Australia.
authors.forename: James G
authors.initials: JG
authors.lastname: Burchfield
authors.forename: Jinling
authors.initials: J
authors.lastname: Lu
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Fazakerley
authors.forename: Shi-Xiong
authors.initials: SX
authors.lastname: Tan
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Ng
authors.forename: Katarina
authors.initials: K
authors.lastname: Mele
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Buckley
authors.forename: Weiping
authors.initials: W
authors.lastname: Han
authors.forename: William E
authors.initials: WE
authors.lastname: Hughes
authors.forename: David E
authors.initials: DE
authors.lastname: James
mesh_terms: D016475:3T3 Cells; D000818:Animals; D051275:Glucose Transporter Type 4 / Q000235:genetics / Q000378:metabolism*; D057166:High-Throughput Screening Assays; D007328:Insulin / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008856:Microscopy, Fluorescence; D021381:Protein Transport
chemical_list: D051275:Glucose Transporter Type 4; D007328:Insulin; D008164:Luminescent Proteins; C495693:Slc2a4 protein, mouse