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    Cells Tissues Organs. 2009;189(1-4):207-11. Epub 2008 Aug 14.

    Immunogold labeling of amelogenin in developing porcine enamel revealed by field emission scanning electron microscopy.

    Du C, Fan D, Sun Z, Fan Y, Lakshminarayanan R, Moradian-Oldak J.

    Source

    Center for Craniofacial Molecular Biology, School of Dentistry, University of Southern California, Los Angeles, Calif. 90033, USA.

    Abstract

    The present study describes a method using immunohistochemical labeling in combination with high-resolution imaging (field emission scanning electron microscopy) to investigate the spatial localization of amelogenins on apatite crystallites in developing porcine enamel. Cross-sections of developing enamel tissue from freeze-fractured pig third molar were treated with antiserum against recombinant mouse amelogenin and immunoreactivity confirmed by Western blot analysis. The samples were then treated with the goat anti-rabbit IgG conjugated with 10-nm gold particles. The control samples were treated with the secondary antibody only. The in-lens secondary electrons detector and quadrant back-scattering detector were employed to reveal the high-resolution morphology of enamel structures and gold particle distribution. The immunolabeling showed a preference of the gold particle localization along the side faces of the ribbon-like apatite crystals. The preferential localization of amelogenin in vivo on enamel crystals strongly supports its direct function in controlling crystal morphology.

    Copyright 2008 S. Karger AG, Basel.

    PMID:
    18701812
    [PubMed - indexed for MEDLINE]
    PMCID: PMC2633245
    Free PMC Article

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