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Sema3A chemorepellant regulates the timing and patterning of dental nerves during development of incisor tooth germ

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Abstract

Semaphorin 3A (Sema3A) axon repellant serves multiple developmental functions. Sema3A mRNAs are expressed in epithelial and mesenchymal components of the developing incisor in a dynamic manner. Here, we investigate the functions of Sema3A during development of incisors using Sema3A-deficient mice. We analyze histomorphogenesis and innervation of mandibular incisors using immunohistochemistry as well as computed tomography and thick tissue confocal imaging. Whereas no apparent disturbances in histomorphogenesis or hard tissue formation of Sema3A −/− incisors were observed, nerve fibers were prematurely seen in the presumptive dental mesenchyme of the bud stage Sema3A −/− tooth germ. Later, nerves were ectopically present in the Sema3A −/− dental papilla mesenchyme during the cap and bell stages, whereas in the Sema3A +/+ mice the first nerve fibers were seen in the pulp after the onset of dental hard tissue formation. However, no apparent topographic differences in innervation pattern or nerve fasciculation were seen inside the pulp between postnatal and adult Sema3A +/+ or Sema3A −/− incisors. In contrast, an abnormally large number of nerves and arborizations were observed in the Sema3A −/− developing dental follicle target field and periodontium and, unlike in the wild-type mice, nerve fibers were abundant in the labial periodontium. Of note, the observed defects appeared to be mostly corrected in the adult incisors. The expressions of Ngf and Gdnf neurotrophins and their receptors were not altered in the Sema3A −/− postnatal incisor or trigeminal ganglion, respectively. Thus, Sema3A is an essential, locally produced chemorepellant, which by creating mesenchymal exclusion areas, regulates the timing and patterning of the dental nerves during the development of incisor tooth germ.

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Acknowledgments

Kjellfrid Haukanes is acknowledged for skilful technical assistance. We thank the staff of the animal facility for careful mouse husbandry. Confocal imaging and computed tomography were performed at the Molecular Imaging Centre (Fuge, Norwegian Research Council), University of Bergen. We thank Cecilie Brekke Rygh for help with the computed tomography. We also thank Stein Atle Lie (Department of Clinical Dentistry, University of Bergen) for help in statistical analysis. This study has been supported by grants from the Norwegian Cancer Society, L. Meltzer’s foundation and Norsk Dental Depot.

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The authors declare no conflict of interest.

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Authors and Affiliations

  1. Craniofacial Developmental Biology Group, Department of Biomedicine, University of Bergen, Jonas Lies vei 91, 5009, Bergen, Norway

    Anjana Shrestha, Kyaw Moe, Keijo Luukko & Paivi Kettunen

  2. Section of Orthodontics, Department of Clinical Dentistry, University of Bergen, Årstadveien 17, 5009, Bergen, Norway

    Keijo Luukko

  3. Department of Biochemistry, Cancer Research Institute, Sapporo Medical University School of Medicine, S-1, W-17, Chuo-ku, Sapporo, 060-8556, Japan

    Masahiko Taniguchi

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  1. Anjana Shrestha
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  2. Kyaw Moe
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  3. Keijo Luukko
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  4. Masahiko Taniguchi
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  5. Paivi Kettunen
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Correspondence to Paivi Kettunen.

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Anjana Shrestha and Kyaw Moe contributed equally to the article.

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Immunohistochemical localization of nerve fibers using peripherin antibody in 100-μm-thick tissue sections of PN7 Sema3A +/+ and Sema3A −/− mandibular incisors. Images shown in Fig 6a, b are shown as a movie (MOV 2938 kb)

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Shrestha, A., Moe, K., Luukko, K. et al. Sema3A chemorepellant regulates the timing and patterning of dental nerves during development of incisor tooth germ. Cell Tissue Res 357, 15–29 (2014). https://doi.org/10.1007/s00441-014-1839-3

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