title: Immunogold labeling of amelogenin in developing porcine enamel revealed by field emission scanning electron microscopy.
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.
authors.affiliation: Center for Craniofacial Molecular Biology, School of Dentistry, University of Southern California, Los Angeles, Calif. 90033, USA.
authors.forename: Chang
authors.initials: C
authors.lastname: Du
authors.forename: Daming
authors.initials: D
authors.lastname: Fan
authors.forename: Zhi
authors.initials: Z
authors.lastname: Sun
authors.forename: Yuwei
authors.initials: Y
authors.lastname: Fan
authors.forename: Rajamani
authors.initials: R
authors.lastname: Lakshminarayanan
authors.forename: Janet
authors.initials: J
authors.lastname: Moradian-Oldak
mesh_terms: D053523:Amelogenin / Q000378:metabolism*; D000818:Animals; D003743:Dental Enamel / Q000254:growth & development* / Q000648:ultrastructure*; D005614:Freeze Fracturing; D007150:Immunohistochemistry; D008855:Microscopy, Electron, Scanning; D034421:Sus scrofa / Q000254:growth & development*; D014070:Tooth / Q000166:cytology
chemical_list: D053523:Amelogenin