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Involvement of RhoA/ROCK in insulin secretion of pancreatic β-cells in 3D culture

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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.

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Authors and Affiliations

  1. Stem Cell and Regenerative Medicine Laboratory, Beijing Institute of Transfusion Medicine, Beijing, 100850, China

    Xiaofang Liu, Fang Yan, Hailei Yao, Mingyang Chang, Jinhua Qin, Yunfang Wang & Xuetao Pei

  2. South China Research Center for Stem Cell & Regenerative Medicine, AMMS, Guangzhou, 510005, China

    Fang Yan, Hailei Yao, Mingyang Chang, Jinhua Qin, Yunfang Wang & Xuetao Pei

  3. Department of Obstetrics and Gynecology, General Hospital of Air Force, Beijing, 10014, China

    Xiaofang Liu

  4. Department of Obstetrics and Gynecology, General Hospital of PLA, Beijing, 100039, China

    Yali Li

Authors
  1. Xiaofang Liu
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  2. Fang Yan
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  3. Hailei Yao
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  4. Mingyang Chang
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  5. Jinhua Qin
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  6. Yali Li
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  7. Yunfang Wang
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  8. Xuetao Pei
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Corresponding authors

Correspondence to Yali Li, Yunfang Wang or Xuetao Pei.

Additional information

Xiaofang Liu, Fang Yan, and Hailei Yao contributed equally to this work

This work was supported by the National High Technology Research and Development Program of China (nos. 2013AA020109 to P.X., 2012AA020501 to W.Y.) the National Nature Science Foundation of China (nos. 81170388 and 31370990 to W.Y.).

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Liu, X., Yan, F., Yao, H. et al. Involvement of RhoA/ROCK in insulin secretion of pancreatic β-cells in 3D culture. Cell Tissue Res 358, 359–369 (2014). https://doi.org/10.1007/s00441-014-1961-2

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  • DOI: https://doi.org/10.1007/s00441-014-1961-2

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