CP27 function is necessary for cell survival and differentiation during tooth morphogenesis in organ culture
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CP27 is a novel gene involved in early vertebrate development that features a distinct expression pattern in developing tooth organs. In order to assess CP27 function in tooth organogenesis, we performed CP27 loss and gain of function studies in our tooth organ culture system. Cap stage mouse molar tooth organs were dissected and cultured for 12 days in serumless, chemically defined medium, and selectively treated either with; (i) CP27 derived peptides as a gain of function approach; (ii) antibodies against CP27 as a loss of function approach; and (iii) rabbit serum as a control for the antibody treatment. Following 12 days of culture, tooth organs were fixed, embedded, sectioned, and further analyzed. Morphometric analysis revealed an increase in tooth size in the peptide treated group and an extensive dissociation of the tooth organ in the CP27 antibody treated group. Controls yielded fully-differentiated crown stage tooth organs with enamel and dentin mineralized tissues. Terminal deoxynucleotidyl transferase biotin-dUTP Nick end labeling staining for apoptosis demonstrated a significant increase of apoptotic cells in the CP27 antibody treated group. Immunohistochemistry for 5D4 keratansulfate proteoglycans in conjunction with changes in cell morphology suggested that cells in the CP27 antibody treated group had undergone a change in cell type toward cartilaginous characteristics. We interpret our studies to indicate that CP27 function alterations dramatically affect tooth organ size as well as individual cell fate and survival during tooth organogenesis. © 2002 Elsevier Science B.V. All rights reserved.
author list (cited authors)
Diekwisch, T., & Luan, X.