title: Laminin γ2 knockout mice rescued with the human protein exhibit enamel maturation defects.
abstract: The epithelial ameloblasts are separated from the maturing enamel by an atypical basement membrane (BM) that is enriched in laminin 332 (LM-332). This heterotrimeric protein (α3, ß3 and γ2 chains) provides structural integrity to BMs and influences various epithelial cell processes including cell adhesion and differentiation. Mouse models that lack expression of individual LM-332 chains die shortly after birth. The lethal phenotype of laminin γ2 knockout mice can be rescued by human laminin γ2 (LAMC2) expressed using a doxycycline-inducible (Tet-on) cytokeratin 14 promoter-rtTA. These otherwise normal-looking rescued mice exhibit white spot lesions on incisors. We therefore investigated the effect of rescue with human LAMC2 on enamel maturation and structuring of the atypical BM. The maturation stage enamel organ in transgenic mice was severely altered as compared to wild type controls, a structured BM was no longer discernible, dystrophic matrix appeared in the maturing enamel layer, and there was residual enamel matrix late into the maturation stage. Microtomographic scans revealed excessive wear of occlusal surfaces on molars, chipping of enamel on incisor tips, and hypomineralization of the enamel layer. No structural alterations were observed at other epithelial sites, such as skin, palate and tongue. These results indicate that while this humanized mouse model is capable of rescue in various epithelial tissues, it is unable to sustain structuring of a proper BM at the interface between ameloblasts and maturing enamel. This failure may be related to the atypical composition of the BM in the maturation stage and reaffirms that the atypical BM is essential for enamel maturation.
authors.affiliation: Laboratory for the Study of Calcified Tissues and Biomaterials, Department of Stomatology, Faculty of Dentistry, Université de Montréal, Montréal, Québec, Canada.
authors.forename: Rima M
authors.initials: RM
authors.lastname: Wazen
authors.affiliation: Laboratory for the Study of Calcified Tissues and Biomaterials, Department of Stomatology, Faculty of Dentistry, Université de Montréal, Montréal, Québec, Canada.
authors.forename: Luiz C
authors.initials: LC
authors.lastname: Viegas-Costa
authors.affiliation: Laboratory for the Study of Calcified Tissues and Biomaterials, Department of Stomatology, Faculty of Dentistry, Université de Montréal, Montréal, Québec, Canada; Department of Biochemistry and Molecular Medicine, Université de Montréal, Montréal, Québec, Canada.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Fouillen
authors.affiliation: Shriners Hospital for Children, Montréal, Montréal, Québec, Canada.
authors.forename: Pierre
authors.initials: P
authors.lastname: Moffatt
authors.affiliation: Department of Medicine, Washington University School of Medicine, St. Louis, MO.
authors.forename: Tracy L
authors.initials: TL
authors.lastname: Adair-Kirk
authors.affiliation: Department of Medicine, Washington University School of Medicine, St. Louis, MO.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Senior
authors.affiliation: Laboratory for the Study of Calcified Tissues and Biomaterials, Department of Stomatology, Faculty of Dentistry, Université de Montréal, Montréal, Québec, Canada; Department of Biochemistry and Molecular Medicine, Université de Montréal, Montréal, Québec, Canada. Electronic address: antonio.nanci@umontreal.ca.
authors.forename: Antonio
authors.initials: A
authors.lastname: Nanci
mesh_terms: D000566:Amelogenesis; D000818:Animals; D001485:Basement Membrane / Q000166:cytology / Q000473:pathology*; D002454:Cell Differentiation; D004658:Enamel Organ / Q000166:cytology / Q000648:ultrastructure*; D005804:Genes, Lethal; D006801:Humans; D007180:Incisor; D007797:Laminin / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D055114:X-Ray Microtomography
chemical_list: C400567:LAMC2 protein, human; C498522:Lamc2 protein, mouse; D007797:Laminin
keywords: Amelogenesis; Basement membrane; Enamel maturation; Humanized mouse model; Laminin γ2