Journal: Acta Histochemica et Cytochemica

Article Title: Coexpression of Ang1 and Tie2 in Odontoblasts of Mouse Developing and Mature Teeth—A New Insight into Dentinogenesis

doi: 10.1267/ahc.13043

Figure Lengend Snippet: Distribution of Ang1 and Tie2 in the first molar tooth of the mouse mandible at E17 and E18. ( a ) HE staining of the first molar of mandible at E18. ( b ) A schema of odontoblast differentiation, showing their positional relationships with basement membrane and ameloblasts. ( c, d, e ) Immunohistochemistry for Ang1 in the first molar at E18. ( d ) Higher magnification of the rectangle in ( c ). ( e ) Higher magnification of the rectangle in ( d ). Lower panel is HE staining of the region corresponded to upper one. Ang1 immunosignals in the distal side of odontoblasts (white arrows) and the proximal side of ameloblasts (a black arrow) in addition to the basement membrane (asterisks). Preameloblasts (an arrowhead) are negative for Ang1. ( f ) In situ hybridization for Ang1 mRNA. Positive signals in odontoblasts (a white arrow) and ameloblasts (a black arrow) and negative signals in pre-ameloblasts (an arrowhead). ( g ) Tie2 immunosignals in the central and distal cusps. ( h ) Higher magnification of the rectangle in ( g ). Cellular processes of odontoblasts (arrowheads) are stained positively. Endothelial cells in the pulp horn circumscribing capillaries are Tie2-positive (black arrows). ( i ) Immunostaining for Tie2 of the first molar tooth germ of mandible at E17 as a control. White and black rectangles are magnified at right panels. Undifferentiated mesenchymal cells (arrowheads) are Tie2-negative, but endothelial cells are Tie2-positive (arrows). ( i ) Western blotting of first molar tooth germ of mandible at E18 with anti-Tie2 antibody (left) and anti-phosphorylated Tie2 antibody. All bars=80 µm.

Article Snippet: To retrieve antigens for the anti-Ang1 antibody, specimens were treated with 1 µg/ml proteinase K (Wako Pure Chemical Industries, Osaka, Japan) for 10 min at RT before the blocking procedure.

Techniques: Staining, Immunohistochemistry, In Situ Hybridization, Immunostaining, Western Blot

Journal: Acta Histochemica et Cytochemica

Article Title: Coexpression of Ang1 and Tie2 in Odontoblasts of Mouse Developing and Mature Teeth—A New Insight into Dentinogenesis

doi: 10.1267/ahc.13043

Figure Lengend Snippet: Distribution of Ang1 and Tie2 after birth. ( a ) Ang1 immunosignals in odontoblasts (a white arrow), predentin (a black arrow), ameloblasts (a white arrowhead) at P3. ( b ) Ang1 immunosignals at P8. Right upper panel is higher magnification of the rectangle in left panel. Right lower panel is HE staining of the same field with upper one. Predentin is clearly stained with Ang1-antibody. Hertwig’s epithelial root sheath is Ang1-negative (a black arrowhead). Ang1 immunosignals in the root of the developing tooth at P15. Odontoblasts (i), osteoblasts (ii), cementoblasts (iii) and periodontal ligament (iv) are Ang1-positive. ( d ) Tie2 immunosignals at P3. ( e ) Higher magnification of the rectangle in ( d ). Tie2 signals are detected in odontoblast processes (a white arrow) embedded in the dentin matrix as well as the distal side of odontoblasts (a black arrow). ( f ) In the mandible at P15, Tie2 signals are detected in odontoblasts (i) but not in osteoblasts (ii), cementoblasts (iii), and periodontal ligament (iv). Note that ameloblasts (asterisks in d, e ) are also Tie2-negative. Bar=80 µm.

Article Snippet: To retrieve antigens for the anti-Ang1 antibody, specimens were treated with 1 µg/ml proteinase K (Wako Pure Chemical Industries, Osaka, Japan) for 10 min at RT before the blocking procedure.

Techniques: Staining

Journal: Acta Histochemica et Cytochemica

Article Title: Coexpression of Ang1 and Tie2 in Odontoblasts of Mouse Developing and Mature Teeth—A New Insight into Dentinogenesis

doi: 10.1267/ahc.13043

Figure Lengend Snippet: Double immunofluorescent confocal laser microscopic analysis. ( a ) Distribution of Ang1 (red) and nestin (green) in differentiating odontoblasts (Od) in contact with the basement membrane (BM), facing pre-ameloblasts (Pre Am) at E18. Ang1 is localized in nestin-positive odontoblast. Without antigen retrieved with proteinase K to protect immunosignals for nestin, Ang1 immunosignals in the basement membrane were undetectable. Original magnification, ×60. ( b ) Both Tie2 (green) and nestin (red) are detected in odontoblasts and cells in the pulp horn (a white arrow) at E18. Original magnification, ×20. ( c ) Colocalization of Tie2 (green) and Ang1 (red) in the first molar of the mandible at 8 weeks after birth. Od, a layer of odontoblasts; PreD, predentin; D, dentin; original magnification, ×60. ( d ) Colocalization of Tie2 (green) and Ang1 (red) in odontoblast processes in the first molar of the mandible at 8 weeks after birth. Original magnification, ×100.

Article Snippet: To retrieve antigens for the anti-Ang1 antibody, specimens were treated with 1 µg/ml proteinase K (Wako Pure Chemical Industries, Osaka, Japan) for 10 min at RT before the blocking procedure.

Techniques: