April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
ROCK Inhibition Regulates the Cell Adhesion of Corneal Endothelial Cells
Author Affiliations & Notes
  • Kyoko Kumagai
    Ophthalmology, Kyoto university, Kyoto, Japan
  • Noriko Koizumi
    Biomedical Engineering, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan
  • Naoki Okumura
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Kenta Yamazaki
    Biomedical Engineering, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan
  • Morio Ueno
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Yuji Sakamoto
    Biomedical Engineering, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan
  • Nagahisa Yoshimura
    Ophthalmology, Kyoto university, Kyoto, Japan
  • Junji Hamuro
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Shigeru Kinoshita
    Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan
  • Footnotes
    Commercial Relationships  Kyoko Kumagai, None; Noriko Koizumi, None; Naoki Okumura, None; Kenta Yamazaki, None; Morio Ueno, None; Yuji Sakamoto, None; Nagahisa Yoshimura, None; Junji Hamuro, None; Shigeru Kinoshita, None
  • Footnotes
    Support  The Adaptable and Seamless Technology Transfer Program through Target-driven R & D (grant number: AS2111180G) from the Japan Science and Technology Agency.
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 6465. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Kyoko Kumagai, Noriko Koizumi, Naoki Okumura, Kenta Yamazaki, Morio Ueno, Yuji Sakamoto, Nagahisa Yoshimura, Junji Hamuro, Shigeru Kinoshita; ROCK Inhibition Regulates the Cell Adhesion of Corneal Endothelial Cells. Invest. Ophthalmol. Vis. Sci. 2011;52(14):6465.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose: : We previously reported that the Rho kinase (ROCK) inhibitor Y-27632 promotes cell adhesion of corneal endothelial cells (CECs) and showed that cell injection into the anterior chamber combined with ROCK inhibitor enabled the transplantation of cultivated CECs without a substrate (ARVO, 2009). This current study was conducted to examine how the ROCK signaling regulates cell adhesion of corneal endothelial cells in vitro.

Methods: : CECs of cynomolgus monkeys were cultured. ROCK-I or ROCK-II genes were knocked-down by siRNA and cells were then seeded at a density of 3.0×103/cm2. Random siRNA was used as a control. The numbers of attached cells were evaluated by CellTiter-Glo® Luminescent Cell Viability Assay (Promega Corp., Madison, Wisconsin) after 24 hours. Next, cultured CECs without knockdown by siRNA were seeded with medium containing 10µM of cytochalasin D, a potent inhibitor of actin polymerization. CECs seeded without cytochalasin D were as a control. The numbers of attached cells were evaluated after 24 hours to examine the involvement of actin polymerization in the cell adhesion of CECs.

Results: : The mean numbers of attached cells were 420.3±79.1% in the ROCK-I siRNA group and 115.5±7.0% in the ROCK-II siRNA group as a ratio of the control siRNA group. The CellTiter-Glo® assay revealed that ROCK-I siRNA significantly enhanced cell adhesion of CECs (p<0.01). Inhibition of actin polymerization by cytochalasin D also demonstrated that cell adhesion was significantly enhanced (199.3±18.6% as a ratio of control cells; p<0.01).

Conclusions: : The findings of this study demonstrate that activation of ROCK-I negatively regulates cell adhesion of CECs, possibly via the actin polymerization. We theorize that the regulation of ROCK activity enables cell injection therapy without the use of a substrate for the treatment of corneal endothelial dysfunction.

Keywords: cornea: endothelium • cornea: basic science • cell adhesions/cell junctions 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×