Deletion of epithelial cell-specific Cdc42 leads to enamel hypermaturation in a conditional knockout mouse model

Biochim Biophys Acta Mol Basis Dis. 2018 Aug;1864(8):2623-2632. doi: 10.1016/j.bbadis.2018.04.015. Epub 2018 Apr 21.

Abstract

Recent evidence suggests that GTPases Rho family plays an important role in tooth development; however, the role of Cdc42 in tooth development remains unclear. We aimed to investigate the function of Cdc42 in tooth development and amelogenesis. We generated an epithelial cell-specific K5-Cdc42 knockout (KO) mouse to evaluate post-eruption dental phenotypes using a K5-Cre driver line. This model overcomes the previously reported perinatal lethality. Tooth phenotypes were analyzed by micro X-ray, micro-computed tomography (CT), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), wear rate, shear strength, and a microhardness test. Enamel matrix protein expression was determined by immunohistochemistry. KO mice displayed a hypomaturation phenotype, including incisors that lacked yellow pigmentation and were abnormally white, rapid attrition of molars following eruption, and decreased micro-hardness and shearing strength. Micro-CT data revealed that of incisor and molar enamel volumes were smaller in the KO than in wild-type (WT) mice. SEM analysis showed that the enamel prism structure was disordered. In addition, HE staining indicated a remarkable difference in the ameloblast morphology and function between KO and WT mice, and immunohistochemistry showed increased expression of amelogenin, ameloblastin, matrix metallopeptidase 20, kallikrein-related peptidase 4 and amelotin in the KO mice teeth. Our results suggest epithelium cell-specific Cdc42 deletion leads to tooth hypomaturation and transformation of the enamel prism structure that is likely due to altered ameloblast morphology and the secretion of enamel matrix proteins and proteases. This is the first in vivo evidence suggesting that Cdc42 is essential for proper tooth development and amelogenesis.

Keywords: Cell division cycle 42; Enamel development; Epithelial-specific knockout; Hypomaturation; Morphogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amelogenesis
  • Animals
  • Dental Enamel / metabolism*
  • Dental Enamel / pathology
  • Epithelial Cells / metabolism*
  • Epithelial Cells / pathology
  • Gene Deletion*
  • Incisor / diagnostic imaging
  • Incisor / metabolism*
  • Incisor / pathology
  • Mice
  • Mice, Knockout
  • Molar / diagnostic imaging
  • Molar / metabolism*
  • Molar / pathology
  • X-Ray Microtomography
  • cdc42 GTP-Binding Protein / genetics*
  • cdc42 GTP-Binding Protein / metabolism

Substances

  • Cdc42 protein, mouse
  • cdc42 GTP-Binding Protein