Visualization of the localization of phospholipids in developing rat teeth by matrix-assisted laser desorption/ionization imaging mass spectrometry

  • SASANO Yasuyuki
    Craniofacial Development and Tissue Biology, Tohoku University Graduate School of Dentistry,
  • KONNO Alu
    Department of Optical Imaging, Institute for Medical Photonics Research, Preeminent Medical Photonics Education & Research Center, Hamamatsu University School of Medicine, Department of Virology and Parasitology, Hamamatsu University School of Medicine,
  • NAKAMURA Megumi
    Craniofacial Development and Tissue Biology, Tohoku University Graduate School of Dentistry,
  • HENMI Akiko
    Craniofacial Development and Tissue Biology, Tohoku University Graduate School of Dentistry,
  • MAYANAGI Miyuki
    Craniofacial Development and Tissue Biology, Tohoku University Graduate School of Dentistry,
  • YANG Mu-Chen
    Craniofacial Development and Tissue Biology, Tohoku University Graduate School of Dentistry,
  • YAO Ikuko
    Department of Optical Imaging, Institute for Medical Photonics Research, Preeminent Medical Photonics Education & Research Center, Hamamatsu University School of Medicine, International Mass Imaging Center, Hamamatsu University School of Medicine, Biomedical Engineering Major, Department of Biomedical Sciences, School of Biological and Environmental Sciences, Kwansei Gakuin University,

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Description

<p>Matrix-assisted laser desorption/ionization imaging mass spectrometry (IMS) is used to comprehensively visualize the spatial distribution of numerous biomolecules. The present study was designed to investigate the distribution of phospholipids in developing rat teeth by IMS to identify the characteristic phospholipid molecules for tooth development, and to evaluate the suitability of tissue preparation methods. Rats at postnatal day 3 were euthanized, and the resected head specimens were either fixed or not fixed with 4% paraformaldehyde (PFA), and decalcified or not decalcified in 10% ethylenediaminetetraacetic acid (EDTA) before being frozen. Subsequently, sections were prepared and mounted on glass slides coated with indium tin oxide, and analyzed by IMS. The mass spectra showed the highest peaks around m/z 706, 732, and 734 in the region of interest. Characteristic localization of signals in the tooth buds was seen around m/z 706 and 732, and a database search indicated that the corresponding molecules were phosphatidylcholines. The signals were localized to the dental papillae and enamel epithelia in the tooth buds. The PFA-fixed specimens with or without EDTA decalcification showed preserved IMS signals, while the non-fixed specimens showed fewer signals. Thus, PFA fixation with EDTA decalcification appears to be suitable for IMS analysis of calcified tissues.</p>

Journal

  • Biomedical Research

    Biomedical Research 44 (4), 173-179, 2023-08-03

    Biomedical Research Press

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