Cytoskeletal dependence of insulin granule movement dynamics in INS-1 beta-cells in response to glucose

For pancreatic β-cells to secrete insulin in response to elevated blood glucose, insulin granules retained within the subplasmalemmal space must be transported to sites of secretion on the plasma membrane. Using a combination of super-resolution STORM imaging and live cell TIRF microscopy we investi...

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Bibliographic Details
Published in:PloS one 2014-10, Vol.9 (10), p.e109082-e109082
Main Authors: Heaslip, Aoife T, Nelson, Shane R, Lombardo, Andrew T, Beck Previs, Samantha, Armstrong, Jessica, Warshaw, David M
Format: Article
Language:English
Subjects:
Actin
Animals
Beta cells
Biology and Life Sciences
Biophysics
Blood glucose
Cell Line
Cytoplasmic Granules - drug effects
Cytoplasmic Granules - metabolism
Cytoskeleton
Cytoskeleton - drug effects
Cytoskeleton - metabolism
Exocytosis - drug effects
Exocytosis - physiology
Glucose
Glucose - pharmacology
Granular materials
Health sciences
Hyperglycemia
Image resolution
Insulin
Insulin - metabolism
Insulin secretion
Insulin-Secreting Cells - drug effects
Insulin-Secreting Cells - metabolism
Microscopy
Microtubules - metabolism
Pancreas
Pancreatic beta cells
Pharmacology
Physiology
Polymerization
Quantum dots
Rats
Secretion
Secretory Vesicles - drug effects
Secretory Vesicles - metabolism
Transport
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Summary:For pancreatic β-cells to secrete insulin in response to elevated blood glucose, insulin granules retained within the subplasmalemmal space must be transported to sites of secretion on the plasma membrane. Using a combination of super-resolution STORM imaging and live cell TIRF microscopy we investigate how the organization and dynamics of the actin and microtubule cytoskeletons in INS-1 β-cells contribute to this process. GFP-labeled insulin granules display 3 different modes of motion (stationary, diffusive-like, and directed). Diffusive-like motion dominates in basal, low glucose conditions. Upon glucose stimulation no gross rearrangement of the actin cytoskeleton is observed but there are increases in the 1) rate of microtubule polymerization; 2) rate of diffusive-like motion; and 3) proportion of granules undergoing microtubule-based directed motion. By pharmacologically perturbing the actin and microtubule cytoskeletons, we determine that microtubule-dependent granule transport occurs within the subplasmalemmal space and that the actin cytoskeleton limits this transport in basal conditions, when insulin secretion needs to be inhibited.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0109082