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Deficient Cell Proliferation in Palatal Shelf Mesenchyme of CL/Fr Mouse Embryos

¹ Section of Pediatric Dentistry, Division of Oral Health, Growth and Development, Faculty of Dental Science, Kyushu University, 3-1-1 Maidashi Higashi-ku, Fukuoka 812-8582, Japan; ² School of Dentistry, Kyushu University, Fukuoka, Japan; and ³ Department of Pathology, Nippon Dental University;

View larger version (102K): [in this window] [in a new window]   Figure 1. Procedures for palatal organ culture in CL/Fr bilateral cleft lip (A,C,E) and CL/Fr normal (B,D,F) embryos. (A) Head of a CL/Fr-BCL embryo; (B) head of a CL/Fr-N embryo; (C) palate of a CL/Fr-BCL embryo with mandible removed; (D) palate of a CL/Fr-N embryo with the mandible removed, showing normal lip closure; (E) transverse section of a CL/Fr-BCL palatal shelf after 48 hrs of culture; and (F) transverse section of a CL/Fr-N palatal shelf after 48 hrs of culture. In (A) and (C), a bilateral cleft lip is indicated by arrows. In (C) and (D), the dotted lines indicate the incision line. In (E) and (F), both palates show epithelial disappearance and mesenchymal fusion (arrows). Scale bars = 1 mm (A,B,C,D) and 500 µm (E,F).

View larger version (122K): [in this window] [in a new window]   Figure 2. Transverse sections of palatal shelves stained by the BrdU method for quantitative assessment of cell proliferation. Many positive cells are observed in all palatal mesenchymal and epithelial regions in the C57BL (B), CL/Fr-BCL (C), and CL/Fr-N (D) embryos compared with the eosin background in the BrdU-free control section (A). The radius of the arch is 200 µm and indicates the area for counting positive cells. *Medial tip of the shelf; scale bar = 100 µm.

View larger version (32K): [in this window] [in a new window]   Figure 3. Mean values for BrdU-positive cells for left- and right-side palatal shelves of C57BL, CL/Fr bilateral cleft lip, and CL/Fr normal embryos. (A) The number of positive cells in the epithelium; (B) the ratio of positive to total number of cells in the epithelium; (C) the number of positive cells in the mesenchyme; and (D) the ratio of positive to total number of cells in the mesenchyme. Black boxes represent the left-side and shaded boxes represent the right-side palatal shelves. Error bars show standard deviation. Number in box shows each sample size. **P < 0.01; *P < 0.05.

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