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Molecular expression of Mg2+ regulator TRPM7 and CNNM4 in rat odontoblasts
Affiliation:1. Department of Brain and Cognitive Sciences, College of Natural Sciences, Seoul National University, Seoul, Republic of Korea;2. Dental Research Institute and Department of Neurobiology & Physiology, School of Dentistry, Seoul National University, Seoul, Republic of Korea;1. Department of Orthodontics, Department of Dentistry, Taipei Medical University Hospital, Taipei, Taiwan;2. Graduate Institute of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei, Taiwan;3. Department of Anatomy, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan;1. Department of Orthodontics, Kermanshah University of Medical Sciences, Kermanshah, Iran;2. Department of Oral and Maxillofacial Medicine, School of Dentistry, Kermanshah University of Medical Sciences, Kermanshah, Iran;3. Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran;4. Department of Endodontics, School of Dentistry, Kermanshah University of Medical Sciences, Kermanshah, Iran;5. Students Research Committee, Kermanshah University of Medical Sciences, Kermanshah, Iran;1. Department of Clinical Pharmacy, School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, 211198, Jiangsu, PR China;2. Department of Geriatric Cardiology, Chinese PLA general hospital, Beijing, 100853, PR China;3. Department of Cardiology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, 211198, Jiangsu, PR China
Abstract:
ObjectiveMagnesium, the second most abundant cation in cellular fluid, is critical for mineralization of hard tissues. Among the molecules involved in cellular Mg2+ homeostasis, functional impairment of Mg2+ permeable ion channel TRPM7 or Mg2+ transporter CNNM4 have been found to result in severe hypomineralization of the enamel and dentin. However, molecular expressions of TRPM7, CNNM4 and their respective homologues have not been fully investigated in adult odontoblasts.DesignExpressions of TRPM6, TRPM7, CNNM1, CNNM2, CNNM3, CNNM4 were screened in acutely dissociated rat odontoblasts by single cell RT-PCR. Among these candidates, expression levels of TRPM7 and CNNM4 were compared along the odontoblast layer by immunohistochemical analysis. Finally, the coexpression pattern of TRPM7 and CNNM4 in subcellular regions was examined by immunocytochemical analysis.ResultsScRT-PCR revealed high expression rate of TRPM7 and CNNM4 in odontoblasts, with CNNM4 detected almost exclusively in TRPM7-positive odontoblasts. However, CNNM2 and CNNM3 were detected in only a small population of odontoblasts, and TRPM6 and CNNM1 were not detected even in the pulp tissue. Immunohistochemical analysis revealed higher CNNM4 expression in the apical odontoblast layer than the coronal area, in contrast to the ubiquitous expression of TRPM7. Lastly, immunocytochemical analysis revealed colocalization of CNNM4 with TRPM7 in the odontoblastic process.ConclusionsCNNM4 and TRPM7 may serve as main Mg2+ regulators in odontoblasts, possibly with selective involvement of CNNM4 in apical dentin formation or mineralization. Colocalization of TRPM7 and CNNM4 in the odontoblastic process suggest functional coupling of these two molecules to maintain Mg2+ homeostasis.
Keywords:Odontoblast(s)  Pulp biology  Magnesium ion  TRP channels  CNNM transporters  Dentin
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