Glucose-stimulated insulin secretion is coupled to the interaction of actin with the t-SNARE (target membrane soluble N-ethylmaleimide-sensitive factor attachment protein receptor protein) complex

Debbie C. Thurmond, Carmen Gonelle-Gispert, Megumi Furukawa, Philippe A. Halban, Jeffrey E. Pessin

Research output: Contribution to journalArticle

125 Citations (Scopus)

Abstract

The actin monomer sequestering agent latrunculin B depolymerized β-cell cortical actin, which resulted in increased glucose-stimulated insulin secretion in both cultured MIN6 β-cells and isolated rat islet cells. In perifused islets, latrunculin B treatment increased both first- and second-phase glucose-stimulated insulin secretion without any significant effect on total insulin content. This increase in secretion was independent of calcium regulation because latrunculin B also potentiated calcium-stimulated insulin secretion in permeabilized MIN6 cells. Confocal immunofluorescent microscopy revealed a redistribution of insulin granules to the cell periphery in response to glucose or latrunculin B, which correlated with a reduction in phalloidin staining of cortical actin. Moreover, the t-SNARE [target membrane soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptor] proteins Syntaxin 1 and SNAP-25 coimmunoprecipitated polymerized actin from unstimulated MIN6 cells. Glucose stimulation transiently decreased the amount of actin coimmunoprecipitated with Syntaxin 1 and SNAP-25, and latrunculin B treatment fully ablated the coimmunoprecipitation. In contrast, the actin stabilizing agent jasplakinolide increased the amount of actin coimmunoprecipitated with the t-SNARE complex and prevented its dissociation upon glucose stimulation. These data suggest a mechanism whereby glucose modulates β-cell cortical actin organization and disrupts the interaction of polymerized actin with the plasma membrane t-SNARE complex at a distal regulatory step in the exocytosis of insulin granules.

Original languageEnglish
Pages (from-to)732-742
Number of pages11
JournalMolecular Endocrinology
Volume17
Issue number4
DOIs
StatePublished - Apr 1 2003

Fingerprint

SNARE Proteins
Actins
Membrane Proteins
Insulin
Glucose
Proteins
Syntaxin 1
jasplakinolide
Sequestering Agents
Calcium
Phalloidine
Excipients
Exocytosis
Islets of Langerhans
Confocal Microscopy
Cultured Cells
Cell Membrane
latrunculin B
Staining and Labeling

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Glucose-stimulated insulin secretion is coupled to the interaction of actin with the t-SNARE (target membrane soluble N-ethylmaleimide-sensitive factor attachment protein receptor protein) complex. / Thurmond, Debbie C.; Gonelle-Gispert, Carmen; Furukawa, Megumi; Halban, Philippe A.; Pessin, Jeffrey E.

In: Molecular Endocrinology, Vol. 17, No. 4, 01.04.2003, p. 732-742.

Research output: Contribution to journalArticle

Thurmond, Debbie C. ; Gonelle-Gispert, Carmen ; Furukawa, Megumi ; Halban, Philippe A. ; Pessin, Jeffrey E. / Glucose-stimulated insulin secretion is coupled to the interaction of actin with the t-SNARE (target membrane soluble N-ethylmaleimide-sensitive factor attachment protein receptor protein) complex. In: Molecular Endocrinology. 2003 ; Vol. 17, No. 4. pp. 732-742.
@article{270d2da1fc4b4394a63f000cc5772f18,
title = "Glucose-stimulated insulin secretion is coupled to the interaction of actin with the t-SNARE (target membrane soluble N-ethylmaleimide-sensitive factor attachment protein receptor protein) complex",
abstract = "The actin monomer sequestering agent latrunculin B depolymerized β-cell cortical actin, which resulted in increased glucose-stimulated insulin secretion in both cultured MIN6 β-cells and isolated rat islet cells. In perifused islets, latrunculin B treatment increased both first- and second-phase glucose-stimulated insulin secretion without any significant effect on total insulin content. This increase in secretion was independent of calcium regulation because latrunculin B also potentiated calcium-stimulated insulin secretion in permeabilized MIN6 cells. Confocal immunofluorescent microscopy revealed a redistribution of insulin granules to the cell periphery in response to glucose or latrunculin B, which correlated with a reduction in phalloidin staining of cortical actin. Moreover, the t-SNARE [target membrane soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptor] proteins Syntaxin 1 and SNAP-25 coimmunoprecipitated polymerized actin from unstimulated MIN6 cells. Glucose stimulation transiently decreased the amount of actin coimmunoprecipitated with Syntaxin 1 and SNAP-25, and latrunculin B treatment fully ablated the coimmunoprecipitation. In contrast, the actin stabilizing agent jasplakinolide increased the amount of actin coimmunoprecipitated with the t-SNARE complex and prevented its dissociation upon glucose stimulation. These data suggest a mechanism whereby glucose modulates β-cell cortical actin organization and disrupts the interaction of polymerized actin with the plasma membrane t-SNARE complex at a distal regulatory step in the exocytosis of insulin granules.",
author = "Thurmond, {Debbie C.} and Carmen Gonelle-Gispert and Megumi Furukawa and Halban, {Philippe A.} and Pessin, {Jeffrey E.}",
year = "2003",
month = "4",
day = "1",
doi = "10.1210/me.2002-0333",
language = "English",
volume = "17",
pages = "732--742",
journal = "Molecular Endocrinology",
issn = "0888-8809",
publisher = "The Endocrine Society",
number = "4",

}

TY - JOUR

T1 - Glucose-stimulated insulin secretion is coupled to the interaction of actin with the t-SNARE (target membrane soluble N-ethylmaleimide-sensitive factor attachment protein receptor protein) complex

AU - Thurmond, Debbie C.

AU - Gonelle-Gispert, Carmen

AU - Furukawa, Megumi

AU - Halban, Philippe A.

AU - Pessin, Jeffrey E.

PY - 2003/4/1

Y1 - 2003/4/1

N2 - The actin monomer sequestering agent latrunculin B depolymerized β-cell cortical actin, which resulted in increased glucose-stimulated insulin secretion in both cultured MIN6 β-cells and isolated rat islet cells. In perifused islets, latrunculin B treatment increased both first- and second-phase glucose-stimulated insulin secretion without any significant effect on total insulin content. This increase in secretion was independent of calcium regulation because latrunculin B also potentiated calcium-stimulated insulin secretion in permeabilized MIN6 cells. Confocal immunofluorescent microscopy revealed a redistribution of insulin granules to the cell periphery in response to glucose or latrunculin B, which correlated with a reduction in phalloidin staining of cortical actin. Moreover, the t-SNARE [target membrane soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptor] proteins Syntaxin 1 and SNAP-25 coimmunoprecipitated polymerized actin from unstimulated MIN6 cells. Glucose stimulation transiently decreased the amount of actin coimmunoprecipitated with Syntaxin 1 and SNAP-25, and latrunculin B treatment fully ablated the coimmunoprecipitation. In contrast, the actin stabilizing agent jasplakinolide increased the amount of actin coimmunoprecipitated with the t-SNARE complex and prevented its dissociation upon glucose stimulation. These data suggest a mechanism whereby glucose modulates β-cell cortical actin organization and disrupts the interaction of polymerized actin with the plasma membrane t-SNARE complex at a distal regulatory step in the exocytosis of insulin granules.

AB - The actin monomer sequestering agent latrunculin B depolymerized β-cell cortical actin, which resulted in increased glucose-stimulated insulin secretion in both cultured MIN6 β-cells and isolated rat islet cells. In perifused islets, latrunculin B treatment increased both first- and second-phase glucose-stimulated insulin secretion without any significant effect on total insulin content. This increase in secretion was independent of calcium regulation because latrunculin B also potentiated calcium-stimulated insulin secretion in permeabilized MIN6 cells. Confocal immunofluorescent microscopy revealed a redistribution of insulin granules to the cell periphery in response to glucose or latrunculin B, which correlated with a reduction in phalloidin staining of cortical actin. Moreover, the t-SNARE [target membrane soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptor] proteins Syntaxin 1 and SNAP-25 coimmunoprecipitated polymerized actin from unstimulated MIN6 cells. Glucose stimulation transiently decreased the amount of actin coimmunoprecipitated with Syntaxin 1 and SNAP-25, and latrunculin B treatment fully ablated the coimmunoprecipitation. In contrast, the actin stabilizing agent jasplakinolide increased the amount of actin coimmunoprecipitated with the t-SNARE complex and prevented its dissociation upon glucose stimulation. These data suggest a mechanism whereby glucose modulates β-cell cortical actin organization and disrupts the interaction of polymerized actin with the plasma membrane t-SNARE complex at a distal regulatory step in the exocytosis of insulin granules.

UR - http://www.scopus.com/inward/record.url?scp=0037384072&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037384072&partnerID=8YFLogxK

U2 - 10.1210/me.2002-0333

DO - 10.1210/me.2002-0333

M3 - Article

VL - 17

SP - 732

EP - 742

JO - Molecular Endocrinology

JF - Molecular Endocrinology

SN - 0888-8809

IS - 4

ER -