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RhoE regulates actin cytoskeleton organization in human periodontal ligament cells under mechanical stress
Institution:1. Department of Orthodontics and Dentofacial Orthopedics, Institute of Health Biosciences, The University of Tokushima Graduate School, 3-18-15 Kuramoto-cho, Tokushima 770-8504, Japan;2. Department of Maxillofacial Orthognathics, Tokyo Medical and Dental University Graduate School, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8549, Japan;1. Competence Centre of Periodontal Research, Bernhard Gottlieb School of Dentistry, Medical University of Vienna, Austria;2. Institut Straumann AG, Basel, Switzerland;3. Department of Stomatology, Xuanwu Hospital, Capital Medical University, Beijing, China;4. New York University, College of Dentistry, Department of Periodontology and Implant Dentistry, New York, NY, USA;5. Division of Orthodontics, Bernhard Gottlieb School of Dentistry, Medical University of Vienna, Vienna, Austria;6. Division of Conservative Dentistry, Periodontology and Prophylaxis, Bernhard Gottlieb School of Dentistry, Medical University of Vienna, Vienna, Austria;1. Department of Physiological Psychology, Tokyo Metropolitan Institute of Medical Science, 2-1-6 Kamikitazawa, Setagaya, Tokyo 156-8506, Japan;2. Department of Sensory Science, Faculty of Health Science and Technology, Kawasaki University of Medical Welfare, 288 Matsushima, Kurashiki 701-0193, Japan;3. Division of Systems Neuroscience, Tohoku University Graduate School of Life Sciences, 2-1-1 Katahira, Aoba-ku, Sendai 980-8577, Japan;1. Kobayashi-Maskawa Institute for the Origin of Particles and the Universe (KMI), Nagoya University, Nagoya 464-8602, Japan;2. Institute for Advanced Research, Nagoya University, Nagoya 464-8602, Japan;3. Department of Physics, Nagoya University, Nagoya 464-8602, Japan;4. The University of Tokyo, International Center for Elementary Particle Physics and Department of Physics, 7-3-1 Hongo, Bunkyo-ku, JP - Tokyo 113-0033, Japan;1. Department of Dental Research, The Affiliated Stomatological Hospital of Kunming Medical University, Kunming, Yunnan 650500, PR China;2. National Engineering Laboratory for Oral Regenerative Medicine, Sichuan University, Chengdu 610041, PR China;3. Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, College of Stomatology, Chongqing Medical University, Chongqing 401147, PR China;4. Department of Orthodontics, The Affiliated Stomatological Hospital of Kunming Medical University, Kunming, Yunnan 650031, PR China;5. Department of Oral and Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu 610041, PR China;1. Department of Obstetrics and Gynecology, Nagasaki University School of Medicine, 7-1, Sakamoto 1-choume, Nagasaki-city 8528501, Japan;2. Department of Obstetrics and Gynecology, Sasebo City General Hospital, Sasebo-city 8578511, Japan
Abstract:ObjectivesRhoE and regulator of G-proteins signalling (RGS) 2 were identified as the up-regulated genes in human periodontal ligament (PDL) cells under compression. RhoE belongs to the Rho GTPase family, and RGS2, a novel family of GTPase-activating proteins, turns off the G-protein signalling. Rho family proteins have recently been known to regulate actin cytoskeleton dynamics in various cell types. In this study, we investigated the involvement of RhoE and RGS2 in the regulation of actin filament organization in the PDL cells under mechanical stress.MethodsHuman PDL cells were cultured and subjected to a static compressive force (3.0 g/cm2) for 48 h. To observe changes in the actin cytoskeleton and the expression of RhoE and RGS2 in response to mechanical stress, immunofluorescence analysis was performed. To examine the role of RhoE and RGS2 in actin filament organization, cells were transfected with antisense S-oligonucleotides (ODNs) to RhoE and RGS2.ResultsCompressive force caused a loss and disassembly of actin stress fibres leading to cell spreading. Immunocytochemical study revealed that RhoE and RGS2 expressions were induced by mechanical stress and localized in the perinuclear and in the cell membrane, respectively. The impaired formation of stress fibres caused by compressive forces was recovered by treatment with antisense S-ODN to RhoE to the control levels. However, addition of antisense S-ODN to RGS2 did not affect the stress fibre formation.ConclusionsThese results indicate that the loss and disassembly of stress fibres due to mechanical stress are mediating RhoE signalling, without the exertion of RGS2.
Keywords:RhoE  Periodontal ligament cells  Mechanical stress
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