Distinct Distribution and Localization of Rho-kinase in Mouse Epithelial, Muscle and Neural Tissues

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  • Iizuka Michiro
    Department of Neural Regeneration and Cell Communication, Mie University Graduate School of Medicine Department of Cell Pharmacology, Nagoya University Graduate School of Medicine
  • Kimura Kazushi
    Department of Neural Regeneration and Cell Communication, Mie University Graduate School of Medicine
  • Wang Shujie
    Department of Neural Regeneration and Cell Communication, Mie University Graduate School of Medicine
  • Kato Katsuhiro
    Department of Cell Pharmacology, Nagoya University Graduate School of Medicine Department of Cardiology, Nagoya University Graduate School of Medicine
  • Amano Mutsuki
    Department of Cell Pharmacology, Nagoya University Graduate School of Medicine
  • Kaibuchi Kozo
    Department of Cell Pharmacology, Nagoya University Graduate School of Medicine CREST, Japan Science and Technology Agency
  • Mizoguchi Akira
    Department of Neural Regeneration and Cell Communication, Mie University Graduate School of Medicine

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The small GTP-binding protein Rho plays a crucial role in a wide variety of cellular functions through various effector proteins. Rho-kinase is a key effector protein of Rho, which is composed of two isoforms, ROCK1 and ROCK2. To clarify the site of action of ROCK1 and ROCK2, we performed immunofluorescence and immunoelectron microscopic analyses using isoform-specific antibodies in mouse tissues. In the large and small intestines, ROCK1 immunoreactivity was predominantly identified in epithelial cells, and ROCK2 immunoreactivity was negligible. In these epithelial cells, ROCK1 immunoreactivity was distributed on plasma membranes, while ROCK1 immunogold signals were localized at cell-cell contacts and cell adhesion sites, especially at the adherens junctions at the ultrastructural level. In the bladder epithelium, however, ROCK1 and ROCK2 signals were identified at intermediate filaments, and ROCK2 signals were also observed in nuclei. In the three types of muscular cells—smooth, cardiac, and skeletal muscle cells—ROCK1 and ROCK2 also showed differential distribution. ROCK1 signals were localized at actin filaments, plasma membranes, and vesicles near plasma membranes in smooth muscle cells; at the lysosomes in skeletal muscle cells; and were undetectable in cardiac muscle cells. ROCK2 signals were localized at actin filaments and centrosomes in smooth muscle cells, at intercalated discs in cardiac muscle cells, and at Z-discs and sarcoplasmic reticulum in skeletal muscle cells. In the brain, ROCK1 immunoreactivity was distributed in glia, whereas ROCK2 immunoreactivity was observed in neurons. These results indicate that the two isoforms of Rho-kinase distribute differentially to accomplish their specific functions.

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