Calbindin-D 28K Localization in Rat Molars During Odontogenesis

Abstract

Specific antiserum raised against Calbindin-D_28K, a vitamin D-dependent calcium-binding protein (CaBP) isolated from chick intestine, was used for localization of the protein in developing rat molars. Previously, CaBP had been localized in specific cells associated with the continuously erupting rat incisor: late pre-secretory ameloblasts, secretory and maturation zone ameloblasts, stratum intermedium cells adjacent to ameloblasts in the late zone of enamel secretion, and papillary cells underlying maturation zone ameloblasts. In this study, the peroxidase anti-peroxidase technique was used for localization of CaBP in histological sections of rat mandibles from 18-day-old rat embryos through 20-day-old neonates. CaBP was not detected in any cells of the enamel organ, dental papilla, or dental sac during early odontogenesis from the dental lamina stage through the advanced bell stage. The protein first appeared in secretory ameloblasts which were situated opposite odontoblasts with newly secreted dentin. CaBP was present in the cytoplasm of more mature ameloblasts, but not in less mature ameloblasts. Some stratum intermedium cells subjacent to well-developed secretory and maturation zone ameloblasts also contained CaBP. The protein was not detected in odontoblasts, pulp cells, or other cells associated with the developing molars. It was also absent from the demineralized enamel and dentin matrix. In developing rat molars, the time-course of appearance of CaBP, a protein dependent for its synthesis on the vitamin D endocrine system in other organ systems, suggests a potential direct role of this hormonal system in enamel mineralization.

Get full access to this article

View all access options for this article.

References

Baimbridge, K.G. and Parkes, C.O. (1981): Vitamin D-dependent Calcium Binding Protein in the Chick Brain, Cell Calcium 2:65-76.

Bishop, C.W. ; Kendrick, N.C. ; and DeLuca, H.F. (1983): Induction of Calcium Binding Protein Before 1,25-Dihydroxyvitamin D3 Stimulation of Duodenal Calcium Uptake , J Biol Chem 258:1305-1310.

Christakos, S. ; Friedlander, E.J. ; Frandsen, B.R. ; and Norman, A.W. (1979): Studies on the Mode of Action of Calciferol. XIII. Development of a Radioimmunoassay for Vitamin D-dependent Chick Intestinal Calcium-binding Protein and Tissue Distribution, Endocrinol 104:1495-1503.

Corradino, R.A. ; Wasserman, R.H. ; Pubols, M.H. ; and Chang, S.I. (1968): Vitamin D3 Induction of a Calcium-binding Protein in the Uterus of the Laying Hen, Arch Biochem Biophys 125:378-380.

Kallfelz, F.A. ; Taylor, A.N. ; and Wasserman, R.H. (1967): Vitamin D-induced Calcium-binding Factor in Rat Intestinal Mucosa, Proc Soc Exp Biol Med 125:54-58.

Kretsinger, R.H. ; Mann, J.E. ; and Simmons, J.G. (1982): Model of Facilitated Diffusion of Calcium by the Intestinal Calcium Binding Protein. In: Vitamin D, Chemical, Biochemical and Clinical Endocrinology, A.W. Norman , K. Schaefer , D.v. Herrath , and H.G. Grigoleit , Eds., Berlin: Walter de Gruyter , pp. 233-248.

Norman, A.W. ; Roth, J. ; and Orci, L. (1982): The Vitamin D Endocrine System: Steroid Metabolism, Hormone Receptors, and Biological Response (Calcium Binding Proteins), Endocrine Revs 3:331-366.

Sternberger, L.A. ; Hardy, P.H. ; Cuculis, J.J. ; and Meyer, H.G. (1970): The Unlabeled Antibody-Enzyme Method of Immunohistochemistry, Preparation and Properties of Soluble Antigen-Antibody Complex (Horseradish Peroxidase-Anti-Horseradish Peroxidase) and Its Use in Identification of Spirochetes , J Histochem Cytochem 18:315-333.

Taylor, A.N. (1974): Chick Brain Calcium-binding Protein: Comparison with Intestinal Vitamin D-induced Calcium-binding Protein, Arch Biochem Biophys 161:100-108.

10.

Taylor, A.N. (1981): Immunochemical Localization of the Vitamin D-induced Calcium-binding Protein: Relocation of Antigen During Frozen Section Processing , J Histochem Cytochem 29 :65-73.

11.

Taylor, A.N. (1983): Intestinal Vitamin D-induced Calcium-binding Protein: Time-course of Immunocytological Localization Following 1,25-Dihydroxyvitamin D3, J Histochem Cytochem 31 :426-432.

12.

Taylor, A.N. (1984): Tooth Formation and the 28,000-Dalton Vitamin D-dependent Calcium-binding Protein: An Immunocytochemical Study, J Histochem Cytochem 32:159-164.

13.

Taylor, A.N. ; Gleason, W.A. ; Langman, C.B. ; and Bour-Deau, J.E. (1986): Sensitivity of Ameloblasts and Intestinal Cells to the Vitamin D Endocrine System, J Dent Res 65:753, Abst. No. 248 (IADR).

14.

Taylor, A.N. ; Gleason, W.A. ; and Lankford, G.L. (1984): Rat Intestinal Vitamin D-dependent Calcium-binding Protein: Immunocytochemical Localization in Incisor Ameloblasts, J Dent Res 63:94-97.

15.

Taylor, A.N. ; Gleason, W.A. ; and Lankford, G.L. (1984): Immunocytochemical Localization of Rat Intestinal Vitamin D-dependent Calcium-binding Protein, J Histochem Cytochem 32 :153-158.

16.

Taylor, A.N. ; Inpanbutr, N. ; and Deftos, L.J. (1987): Localization of Calbindin-D28K in Calcitonin Containing Cells of Chick Ultimobranchial Glands, Anat Rec (in press).

17.

Taylor, A.N. and Wasserman, R.H. (1967): Vitamin D-induced Calcium-binding Protein: Partial Purification, Electrophoretic Visualization, and Tissue Distribution, Arch Biochem Biophys 119:536-540.

18.

Taylor, A.N. and Wasserman, R.H. (1969): Correlations Between the Vitamin D-induced Calcium Binding Protein and Intestinal Absorption of Calcium, Fed Proc 28:1834-1838.

19.

Wasserman, R.H. ; Corradino, R.A. ; Feher, J. ; and Arm-Brecht, H.J. (1977): Temporal Patterns of Response of the Intestinal Calcium Absorptive System and Related Parameters to 1,25-Dihydroxycholecalciferol . In: Vitamin D Biochemical, Chemical and Clinical Aspects Related to Calcium Metabolism, A.W. Norman, K. Schaefer, J.W. Coburn , H.F. DeLuca, D. Fraser, H.G. Grigoleit , and D.v. Herrath, Eds., Berlin: Walter de Gruyter, pp. 331-340.

20.

Wasserman, R.H. and Fullmer, C.S. (1982): Vitamin D-induced Calcium-binding Protein. In: Calcium and Cell Function. Vol. II, W.Y. Cheung , Ed., New York: Academic Press, pp. 175-216.

21.

Wasserman, R.H. and Taylor, A.N. (1966): Vitamin D3-induced Calcium-binding Protein in Chick Intestinal Tissue, Science 152:791-793.