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Involvement of RhoA/ROCK in insulin secretion of pancreatic β-cells in 3D culture.
- Source :
- Cell & Tissue Research; Nov2014, Vol. 358 Issue 2, p359-369, 11p
- Publication Year :
- 2014
-
Abstract
- Cell-cell contacts and interactions between pancreatic β-cells and/or other cell populations within islets are essential for cell survival, insulin secretion, and functional synchronization. Three-dimensional (3D) culture systems supply the ideal microenvironment for islet-like cluster formation and functional maintenance. However, the underlying mechanisms remain unclear. In this study, mouse insulinoma 6 (MIN6) cells were cultured in a rotating 3D culture system to form islet-like aggregates. Glucose-stimulated insulin secretion (GSIS) and the RhoA/ROCK pathway were investigated. In the 3D-cultured MIN6 cells, more endocrine-specific genes were up-regulated, and GSIS was increased to a greater extent than in cells grown in monolayers. RhoA/ROCK inactivation led to F-actin remodeling in the MIN6 cell aggregates and greater insulin exocytosis. The gap junction protein, connexin 36 (Cx36), was up-regulated in MIN6 cell aggregates and RhoA/ROCK-inactivated monolayer cells. GSIS dramatically decreased when Cx36 was knocked down by short interfering RNA and could not be reversed by RhoA/ROCK inactivation. Thus, the RhoA/ROCK signaling pathway is involved in insulin release through the up-regulation of Cx36 expression in 3D-cultured MIN6 cells. [ABSTRACT FROM AUTHOR]
- Subjects :
- PANCREATIC beta cells
CELL culture
EXOCYTOSIS
GENETIC regulation
LABORATORY mice
Subjects
Details
- Language :
- English
- ISSN :
- 0302766X
- Volume :
- 358
- Issue :
- 2
- Database :
- Complementary Index
- Journal :
- Cell & Tissue Research
- Publication Type :
- Academic Journal
- Accession number :
- 99076438
- Full Text :
- https://doi.org/10.1007/s00441-014-1961-2